320 results for crispr

CRISPR-CasĪ¦ from huge phages is a hypercompact genome editor.

Publication  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems name : CRISPR-CasĪ¦ from huge phages is a hypercompact genome editor. description : CRISPR-Cas systems are found widely in prokaryotes, where they provide adaptive immunity against virus We describe a minimal functional CRISPR-Cas system, comprising a single ~70-kilodalton protein, CasĪ¦, and a CRISPR array, encoded exclusively in the genomes of huge bacteriophages. CasĪ¦ uses a single active site for both CRISPR RNA (crRNA) processing and crRNA-guided DNA cutting to vitro and in human and plant cells with expanded target recognition capabilities relative to other CRISPR-Cas
Pausch P, Al-Shayeb B, Bisom-Rapp E, Tsuchida CA, Li Z, Cress BF, Knott GJ, Jacobsen SE, Banfield JF, Doudna JA
PII: 369/6501/333, PUBMED 32675376, PMC PMC8207990, MID NIHMS1702779, DOI 10.1126/science.abb1400

ABSTRACT: CRISPR-Cas systems are found widely in prokaryotes, where they provide adaptive immunity against virus infection and plasmid transformation. We describe a minimal functional CRISPR-Cas system, comprising a single ~70-kilodalton protein, CasĪ¦, and a CRISPR array, encoded exclusively in the genomes of huge bacteriophages. CasĪ¦ uses a single active site for both CRISPR RNA (crRNA) processing and crRNA-guided DNA cutting to target foreign nucleic acids. This hypercompact system is active in vitro an ...

Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides

Project  [Delivery Systems Initiative]
Matched Fields: name : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02 NIH Report
 
The proposed research is relevant to the public health because genetic and acquired diseases affecting the airways pose major disease and economic burdens. By advancing the delivery of gene editing tools, it may be possible to therapeutically modify the cells lining the airways. This novel strategy has implications for the treatment of both monogenetic and acquired lungs disease, and may have applications for other somatic cell therapies.

Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain

Project  [Delivery Systems Initiative]
Matched Fields: name : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain description : In vivo genome editing using CRISPR/Cas9 is anticipated to be the next wave of therapeutics for various
Gong Shaoqin (Sarah)  Last Updated Date: 2020-10-28 NIH Report
 
In vivo genome editing using CRISPR/Cas9 is anticipated to be the next wave of therapeutics for various major health threats, including neurodegenerative diseases. However, to date, very few Cas9-gRNA ribonucleoprotein in vivo delivery methods have been reported, and delivery to the brain has been particularly challenging. The unique nanocapsules we plan to develop will ultimately enable high efficiency neuron-targeted genome editing in the brain, thereby offering new hope to treat devastating neurodegenerative diseases.

Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain name : Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain. description : Nanocapusules carrying CRISPR Cas9 RNP with guide RNA targeting the stop sequence in the Ai14 transgene experimentName : Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
Gong Shaoqin (Sarah)  Last Updated Date: 2020-10-28
 
Nanocapusules carrying CRISPR Cas9 RNP with guide RNA targeting the stop sequence in the Ai14 transgene are intracerebrally delivered to Ai14 mice and gene editing is measured by gain of tdTomato protein expression.

Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases

Project  [Delivery Systems Initiative]
Matched Fields: name : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Gao Guang-Ping  Last Updated Date: 2020-10-20 NIH Report
 
Efficacy and safety limitations in current gene editing technologies have hindered efforts to treat genetic lung diseases. This proposal seeks to develop and validate a combinatorial delivery approach that uses non-viral and viral vehicles to efficiently transport gene editing tools to disease-relevant cells in the lung. Completion of our work will establish safe and effective delivery vehicles that will guide the design of future gene therapies for genetic disorders.

A bacterial cytidine deaminase toxin enables CRISPR-free mitochondrial base editing.

Publication 
Matched Fields: name : A bacterial cytidine deaminase toxin enables CRISPR-free mitochondrial base editing. description : their use in base editing requires the unwinding of double-stranded DNA (dsDNA)-for example by a CRISPR-Cas9 CRISPR-free DdCBEs enable the precise manipulation of mtDNA, rather than the elimination of mtDNA copies
Raguram A, Liu DR, Mok BY, de Moraes MH, Zeng J, Bosch DE, Kotrys AV, Hsu F, Radey MC, Peterson SB, Mootha VK, Mougous JD
PII: 10.1038/s41586-020-2477-4, PUBMED 32641830, PMC PMC7381381, MID NIHMS1597977, DOI 10.1038/s41586-020-2477-4

ABSTRACT: Bacterial toxins represent a vast reservoir of biochemical diversity that can be repurposed for biomedical applications. Such proteins include a group of predicted interbacterial toxins of the deaminase superfamily, members of which have found application in gene-editing techniques1,2. Because previously described cytidine deaminases operate on single-stranded nucleic acids3, their use in base editing requires the unwinding of double-stranded DNA (dsDNA)-for example by a CRISPR-Cas9 system. Base ...

CHANGE-seq reveals genetic and epigenetic effects on CRISPR-Cas9 genome-wide activity.

Publication  - [In Vitro] [Biological Systems] [Human]
Matched Fields: name : CHANGE-seq reveals genetic and epigenetic effects on CRISPR-Cas9 genome-wide activity. description : Current methods can illuminate the genome-wide activity of CRISPR-Cas9 nucleases, but are not easily
Lazzarotto CR, Malinin NL, Li Y, Zhang R, Yang Y, Lee G, Cowley E, He Y, Lan X, Jividen K, Katta V, Kolmakova NG, Petersen CT, Qi Q, Strelcov E, Maragh S, Krenciute G, Ma J, Cheng Y, Tsai SQ
PII: 10.1038/s41587-020-0555-7, PUBMED 32541958, PMC PMC7652380, MID NIHMS1591991, DOI 10.1038/s41587-020-0555-7

ABSTRACT: Current methods can illuminate the genome-wide activity of CRISPR-Cas9 nucleases, but are not easily scalable to the throughput needed to fully understand the principles that govern Cas9 specificity. Here we describe 'circularization for high-throughput analysis of nuclease genome-wide effects by sequencing' (CHANGE-seq), a scalable, automatable tagmentation-based method for measuring the genome-wide activity of Cas9 in vitro. We applied CHANGE-seq to 110 single guide RNA targets across 13 thera ...

Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia.

Publication  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides name : Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway description : These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas)
Krishnamurthy S, Wohlford-Lenane C, Kandimalla S, Sartre G, Meyerholz DK, ThƩberge V, HallƩe S, DuperrƩ AM, Del'Guidice T, Lepetit-Stoffaes JP, Barbeau X, Guay D, McCray PB
PII: 10.1038/s41467-019-12922-y, PUBMED 31659165, PMC PMC6817825, DOI 10.1038/s41467-019-12922-y

ABSTRACT: The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle ...

Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors

Project  [Delivery Systems Initiative]
Matched Fields: name : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors description : RNA-guided CRISPR genome editing systems promise to revolutionize the treatment of inherited disease. that directs editing is a critical hurdle in the development of clinical applications for engineered CRISPR nucleic acid therapeutics, we have established a framework for complete chemical modification of CRISPR
Sontheimer Erik J  Last Updated Date: 2021-04-15 NIH Report
 
RNA-guided CRISPR genome editing systems promise to revolutionize the treatment of inherited disease. Safe, effective, and target-tissue-specific delivery of the guide RNA that directs editing is a critical hurdle in the development of clinical applications for engineered CRISPR systems. Using strategies validated for the delivery of other categories of nucleic acid therapeutics, we have established a framework for complete chemical modification of CRISPR guides, thereby conferring in vivo stability and effective biodistribution properties. The proposed research will optimize these guides, as well as other editing components, for clinical use.
Show Experiments (11)

Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells
Testing newly chemically modified crRNA and tracrRNA in mTmG mouse embryonic fibroblasts
Testing newly chemically modified crRNA and tracrRNA to activate the TLR1 reporter in human cells
Testing newly chemically modified crRNA and tracrRNA in mouse Hepa 1-6 cells
Testing newly chemically modified crRNA and tracrRNA in mouse Neuro 2A cells
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain
Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center

Self-delivering, chemically modified CRISPR RNAs for AAV co-delivery and genome editing in vivo.

Publication 
Matched Fields: name : Self-delivering, chemically modified CRISPR RNAs for AAV co-delivery and genome editing in vivo. description : Guide RNAs offer programmability for CRISPR-Cas9 genome editing but also add challenges for delivery.
Newby GA, Liu DR, Zhang H, Kelly K, Lee J, Echeverria D, Cooper D, Panwala R, Amrani N, Chen Z, Gaston N, Wagh A, Xie J, Gao G, Wolfe SA, Khvorova A, Watts JK, Sontheimer EJ
PII: 7456042, PUBMED 38033325, PMC PMC10810193, DOI 10.1093/nar/gkad1125

ABSTRACT: Guide RNAs offer programmability for CRISPR-Cas9 genome editing but also add challenges for delivery. Chemical modification, which has been key to the success of oligonucleotide therapeutics, can enhance the stability, distribution, cellular uptake, and safety of nucleic acids. Previously, we engineered heavily and fully modified SpyCas9 crRNA and tracrRNA, which showed enhanced stability and retained activity when delivered to cultured cells in the form of the ribonucleoprotein complex. In this ...

[Validation] Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: name : Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear description : Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice experimentName : Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
Murray Stephen A  Last Updated Date: 2023-05-10
 
Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice

Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems

Project  [Genome Editors]
Matched Fields: name : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems description : Expanding genome editing tools through exploration of new CRISPR-Cas proteins and DNA repair enzymes NARRATIVE Fundamental research on bacterial adaptive immunity uncovered the genome editing properties of CRISPR-Cas We will focus our investigation on newly described CRISPR-Cas systems and DNA-interacting proteins that
Banfield Jillian , Doudna Jennifer A  Last Updated Date: 2020-10-16 NIH Report
 
Expanding genome editing tools through exploration of new CRISPR-Cas proteins and DNA repair enzymes NARRATIVE Fundamental research on bacterial adaptive immunity uncovered the genome editing properties of CRISPR-Cas systems, and it is clear that uncultivated microbes contain more pathways and enzymes that may be useful as genome editing tools. We will combine bioinformatics and biochemistry to identify new DNA- and RNA-associating proteins and will analyze their mechanisms of action. We will focus our investigation on newly described CRISPR-Cas systems and DNA-interacting proteins that occur in conserved genomic context.

[Validation] Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: name : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 description : Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice experimentName : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9
Murray Stephen A  Last Updated Date: 2023-05-10
 
Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice

[Validation] Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: name : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia description : Ribonucleoproteins for CRISPR/Cas9 editing are complexed with amphiphilic peptides for delivery to lung experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
Heaney Jason D  Last Updated Date: 2021-09-07
 
Ribonucleoproteins for CRISPR/Cas9 editing are complexed with amphiphilic peptides for delivery to lung airway epithilia via intranasal instillation into mTmG reporter mice. Editing is detected by production of GFP protein, and green fluorescence in airway linings

[Validation] Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: name : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to description : Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Murray Stephen A  Last Updated Date: 2023-05-10
 
Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice

Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CB promoter)

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: name : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
Asokan Aravind  Last Updated Date: 2021-09-16
 
A dual vector strategy was employed: one delivering a single guide RNA and CB driven SaCas9, and another delivering the second guide RNA and CB driven SaCas9. This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 2e12vg was injected into each mouse (1e12vg each vector mixed 1:1) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.

Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter)

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: name : Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 experimentName : Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
Asokan Aravind  Last Updated Date: 2021-09-16
 
A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:4 ratio of cas9 to guide RNA (1e12vg of CMV Sacas9 vector and 3e12vg of the sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.

Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter)

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: name : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to
Asokan Aravind  Last Updated Date: 2021-09-16
 
A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=3) and AAVcc47 (n=3) by intravenous injection in Ai9 mice. A total dose of 3e12vg was injected into each mouse (1.5e12vg each vector mixed 1:1) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.

Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CMV promoter) and self complementary sgRNA vector.

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: name : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
Asokan Aravind  Last Updated Date: 2021-09-16
 
A dual vector strategy was employed: one self complementary vector delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (single stranded vector). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=4) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:1 ratio of cas9 to guide RNA (2e12vg of CMV Sacas9 vector and 2e12vg of the self complementary sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.

Ai14 gRNA

Guide  - [In Vivo] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
This sgRNA targets the Ai9 and related transgenes at multiple sites

Ai14 mouse

Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
Ai14 mouse has a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus. The att site flanked neo selection cassette has been removed in this strain.

[Validation] Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: description : Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery.
Heaney Jason D  Last Updated Date: 2021-03-30
 
Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery. Detection of editing in non-target tissues.

Ai14 mouse (congenic)

Model System  - [In Vivo] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
Ai14 mouse has a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus. The att site flanked neo selection cassette has been removed in this strain.

Gong_Intracranial Injection Procedure for Mice

Protocol  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
Procedure for intracranial delivery to mouse brain.

Heaney-SATC_McCray-Validation_Intranasal Instillation in Mice Protocol

Protocol  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
Procedure for intranasal instillation of ribonucleoprotein/peptide complex in mice for delivery to the lung.

AB_141607 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: AB_141607 A21202
Invitrogen, Donkey anti-mouse alexafluor 488

D10

Delivery System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia

D237

Delivery System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia

RNP-NC-RVG

Delivery System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
The nanocapsule is a thin glutathione (GSH)-cleavable covalently crosslinked polymer coating around a preassembled ribonucleoprotein (RNP) complex between a Cas9 nuclease and an sgRNA. This nanoparticle has an addition of a RVG peptide YTIWMPENPRPGTPCDIFTNSRGKRASNG which specifically interacts withthe N-acetylecholine receptor (AchR) on neuronal cells, which mediates NP entry

Murray-SATC_Gong-Validation Brain Injection in Mice Protocol

Protocol  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Procedure for brain injection surgical procedure, pre- and post-operative care for mice.

AB_10711040 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: AB_10711040 ab104224Ā 
Abcam, mouse anti-NeuN

SaCas9-Lagor

Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9

[Validation] Independent validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: description : Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing
Heaney Jason D  Last Updated Date: 2023-05-10
 
Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing. Editing detected and quantified in brain by generation of tdTomato fluorescent protein signal from Ai9 reporter mice

SpyCas9 g-loxP2_C9

Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
This sgRNA targets the mTmG transgene

Murray-SATC_Gong-Validaiton_Gong Study Protocol

Protocol  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Procedure for intracranial injection, immunofluorescence and imaging.

AB_2209751 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: Rockland Cat# 600-401-379 AB_2209751
Anti-RFP (RABBIT) Antibody

AAV9-Ai9-sgRNA1-CB-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus

AAVcc47-Ai9-sgRNA2-CB-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus

AAV9_pTR_self comp 2xU6-Ai9 guides

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene

Ai9 mouse

Model System  - [In Vivo] [Small Animal Testing Center (SATC), AAV tropism, Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
Ai9 mouse has a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus.
Show Experiments (11)

Testing gRNA sequence and gRNA scaffold modified in Ai9 mice.
Testing AAV5 for activation of tdTomato in mouse airway
Cre Recombinase dose escalation study in Ai9 mice
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CB promoter)
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter)
Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter)
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CMV promoter) and self complementary sgRNA vector.
Independent validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain
FUS (focused ultrasound) array validation in Ai9 mice
Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cells
AAV Tropism project

Focused Ultrasound-mediated Delivery of Gene-editing Elements to the Brain for Neurodegenerative Disorders

Project  [Delivery Systems Initiative]
Matched Fields: description : Current methodologies of delivering CRISPR-based gene editing elements to the brain are highly inefficient
Leong Kam W  Last Updated Date: 2022-04-15 NIH Report
 
Gene editing may offer a new therapeutic strategy to tackle many neurodegenerative disorders that remain untreatable. Current methodologies of delivering CRISPR-based gene editing elements to the brain are highly inefficient. We propose to develop a noninvasive, efficient approach to achieve gene editing in the brain using focused ultrasound technology.

SpCas9

Genome Editor  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia

sNLS-SpCas9-sNLS

Genome Editor  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
SpCas9 with N- and C-terminal SV40 NLS

RNP-NC-CPP

Delivery System  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
The nanocapsule is a thin glutathione (GSH)-cleavable covalently crosslinked polymer coating around a preassembled ribonucleoprotein (RNP) complex between a Cas9 nuclease and an sgRNA. This nanoparticle has an addition of a cell penetrating peptide (CPP) from the TAT peptide (GRKKRRQRRRPQ) which lacks cell-type specficity

[Validation] Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: description : AAV5 encoding CRISPR/Cas editing machinery were delivered to the lungs of reporter mice by intratracheal
Heaney Jason D  Last Updated Date: 2021-03-30
 
AAV5 encoding CRISPR/Cas editing machinery were delivered to the lungs of reporter mice by intratracheal instillation. After 4 weeks incubation, the mice were dissected and the lungs imaged for the presence of tdTomato fluorescence, indicating successful editing. Editing calculated by dividing the number of tdTomato+ red cells by the number of nuclei in each airway

Podocyte-specific gene editing in human kidney organoids

Experiment  - [In Vitro] [Collaborative Opportunity Fund] [Human]
Matched Fields: description : Intact kidney organoids were transfected with CRISPR ribonucleoprotein complexes with and without molecular
Wilson Ross C. , Freedman Benjamin S  Last Updated Date: 2024-01-16
 
Kidney organoids were derived from a human iPS cell line with Ai9 (tdTomato) fluorescence-on reporter knocked into the AAVS1 safe harbor locus. Intact kidney organoids were transfected with CRISPR ribonucleoprotein complexes with and without molecular targeting agent (MTA) specific for podocytes. Genome editing events were detected by induction of tdTomato from the Ai9 reporter.

In vivo gene editing in dystrophic mouse muscle and muscle stem cells.

Publication 
Matched Fields: description : Delivery by adeno-associated virus (AAV) of clustered regularly interspaced short palindromic repeats (CRISPR AAV-Dmd CRISPR treatment partially recovered muscle functional deficiencies and generated a pool of endogenously
Tabebordbar M, Zhu K, Cheng JKW, Chew WL, Widrick JJ, Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe LH, Church GM, Wagers AJ
PUBMED: 26721686, PMC PMC4924477, MID NIHMS791917, DOI 10.1126/science.aad5177

ABSTRACT: Frame-disrupting mutations in the DMD gene, encoding dystrophin, compromise myofiber integrity and drive muscle deterioration in Duchenne muscular dystrophy (DMD). Removing one or more exons from the mutated transcript can produce an in-frame mRNA and a truncated, but still functional, protein. In this study, we developed and tested a direct gene-editing approach to induce exon deletion and recover dystrophin expression in the mdx mouse model of DMD. Delivery by adeno-associated virus (AAV) of c ...

[Validation] Independent validation of Chaikof delivery platform using virus-like particle (VLP) delivery to the mouse liver

Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: description : Virus-like particles carrying a CRISPR/Cas base editor and a guide RNA targeting the PSCK9 locus were
Heaney Jason D  Last Updated Date: 2023-12-18
 
Virus-like particles carrying a CRISPR/Cas base editor and a guide RNA targeting the PSCK9 locus were injected i.v. into male and female mice. One week after injection, the mice were dissected, and genomic DNA isolated from a panel of organs. Targeted NGS was performed to evaluate the degree of editing at the PCSK9 locus in the liver (primary target), and non-target organs. Two potential off-target editing sites (OT6 and OT7) were also sequenced.

306-S10

Delivery System  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
combinatorial library cationic lipid nanoparticles

Heaney_SATC Tissue Processing, Imaging and Analysis

Protocol  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
Procedure for tissue preparation, imaging and analysis.

Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia.

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
Delivery of GFP via shuttle peptides to mouse airway epithelium via nasal instilation. Delivery efficiency was quantified in large and small airways by counting the number of GFP positive cells divided by the number of DAPI cells.

Gene editing in vitro by various peptide variants delivering Cas12a in NK cells.

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
In Vitro shuttle peptide delivery of Cas12a RNP targeting human NKG2A gene in human NK cells. Gene editing was assessed after 48hr by T7E1 digestion.

Gene editing in vitro by various peptide variants delivering Cas RNPs to primary Human airway epithelia cells.

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
In Vitro shuttle peptide delivery of Cas12a RNPs targeting human CFTR and HPRT genes in human primary airway epithelia. Editing efficiency was assessed after 72hrs by sanger sequencing.

Cre recombinase

Genome Editor  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Cre recombinase delivered by plasmid (see vector details)

FSD168 cyclic

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168d17

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD193

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD236

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD239

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD291

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD306

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD319

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD329

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD330

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD331

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD332

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD341

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD363

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX2

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX3

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

pAAV.pU1a-SpCas9

Vector  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS

3849 45-5'

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Targets endogenous human locus

NKG2A

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
gRNA targeting human NKG2A exon 3

sgAi9L

Guide  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
This sgRNA targets the Ai9 and related transgenes

mTmG mouse

Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
ROSAmT/mGĀ is a cell membrane-targeted, two-color fluorescent Cre-reporter allele. Prior to Cre recombination, cell membrane-localized tdTomato (mT) fluorescence expression is widespread in cells/tissues. Cre recombinase expressing cells (and future cell lineages derived from these cells) have cell membrane-localized EGFP (mG) fluorescence expression replacing the red fluorescence

HEK-293T with Ai9 transient reporter assay

Model System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
HEK-293T cells transfected with an Ai9 inducible transgene reporter plasmid used to test gene editing activity by fluorescence. HEK293T is an epithelial-like cell that was isolated from the kidney of a patient.

NK

Model System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Natural Killer cells. White blood cells;

Peptide Shuttle optimization to deliver Cas12a RNP to human airway epithelia cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
Delivery of Cas12a RNP targeting human CFTR in Primary human epithelia cells. Gene editing efficiency was determined by percentage of NGS reads that showed an indel

Testing AAV5 for activation of tdTomato in mouse airway

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Gao Guang-Ping  Last Updated Date: 2020-10-20
 
AAV2/5 mediated gene editing in the mouse airway was tested by deliverying SpCas9 and guide RNAs targeting the Ai9 transgene in Ai9 transgenic mice. Viral delivery was detected by GFP expression and gene editing quantified by tdTomato activation

Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cells

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Gao Guang-Ping  Last Updated Date: 2021-09-21
 
AAV2/5 mediated gene editing in the mouse airway was tested by deliverying SpCas9 and guide RNAs targeting the Ai9 transgene in Ai9 transgenic mice. Gene editing quantified by tdTomato activation and cell specific markers for club and ciliated cell types.

GFP-NLS

Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Nuclear targeted GFP

FSD112

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD122

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD147

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD159

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168d10

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168D20

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD216

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD222

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD288

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD323

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD367

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57d6

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD67

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD96

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX4

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

S10 Scr.

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

S18

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide

g-loxPbot_C12a

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
This sgRNA targets the Ai9 and related transgenes at two sites

BALB/c mouse

Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
BALB/cJ is a commonly used inbred. Key traits include a susceptibility to developing the demyelinating disease upon infection with Theiler's murine encephalomyelitis virus. The BALB/cJ substrain is susceptible to Listeria, all species of Leishmania, and several species of Trypanosoma, but is resistant to experimental allergic orchitis (EAO).

Lipofectamine 3000

Delivery System  - [In Vitro] [Genome Editors, Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Lipid nanoparticle

CleanCapĀ® Cas9 mRNA (5moU)

Genome Editor  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
SpCas9 mRNA with 2 NLS signals, HA tag and capped using CleanCap. It is polyadenylated, substituted with a modified uridine and optimized for mammalian systems. It mimics a fully processed mature mRNA.

FSD117D1

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Shuttle peptide used to deliver reagents to airway epithelia

Testing newly chemically modified crRNA and tracrRNA in mTmG mouse embryonic fibroblasts

Experiment  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were delivered by electroporation to embryonic fibroblasts harvested from the mTmG reporter mouse. Gene editing was determined by reporter activation.

Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were injected into the intra-striatum of mTmG reporter mice and activation of GFP expression was imaged.

STS204 (DNMT1; Sp_t41:Sp_c20_Dnmt1)

Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets endogenous mouse locus

HEK-293T-disrupted_GFP with MCV-mcherry-Puro

Model System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
HEK293T cells with an integrated reporter for TLR-MCV1 reporter editing. HEK293T is an epithelial-like cell that was isolated from the kidney of a patient.

Testing newly chemically modified crRNA and tracrRNA in mouse Hepa 1-6 cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were delivered by electroporation to mouse Hepa 1-6 cells. Editing activity was determined by Sanger sequencing

STS119 (TLR-MCV1b; Sp_t2:Sp_c20_TLR_MCV1b)

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets traffic light reporter transgene

STS134 (PCSK9b; Sp_t2:Sp_c20_PCSK9b)

Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets endogenous mouse locus

STS159 (mTmG; Sp_t2-PS:Sp_c20_mTmG)

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
This sgRNA targets the mTmG transgene

STS159 (mTmG; Sp_t2:Sp_c0_mTmG)

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
This sgRNA targets the mTmG transgene

STS205 (BACE1; Sp_t41:Sp_c20_Bace1)

Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets endogenous mouse locus

S10

Delivery System  - [In Vivo, In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Mouse, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Shuttle peptide used to deliver reagents to airway epithelia
Show Experiments (12)

Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia.
Peptide Shuttle optimization to deliver Cas9 RNP to human airway epithelia cells
Peptide Shuttle optimization to deliver Cas12a RNP to human airway epithelia cells
Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia
Gene editing in vitro by various peptide variants delivering Cas12a in NK cells.
Gene editing in vitro by various peptide variants delivering Cas RNPs to primary Human airway epithelia cells.
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
Delivery of Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting CFTR locus
Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting B2M locus
Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to rhesus monkey airway epithelial cells cultured at the air liquid interface targeting CCR5 locus.
Inhibitory effect of Cas9 RNP alone on S10- or FSDS315-mediated GFP protein delivery to HeLa cells
Amphiphilic Peptides Deliver Base Editor RNPs to Rhesus Monkey Airway

sg298

Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
This sgRNA targets the Ai9 and related transgenes at multiple sites. 2'-O-Methyl at 3 first and last bases, 3' phosphorothioate bonds between first 3 and last 2 bases

Validating Gene Editing Reporter 14 (GER14) Mouse Model in Heterozygous Blastocysts

Experiment  - [In Vitro] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: description : Zyotes were electroporated with various CRISPR/Cas9 combinations (or Cre) and were cultured 96 hours
Murray Stephen A  Last Updated Date: 2024-02-23
 
WT female mouse and homozygous male mouse containing the Gene Editing Reporter 14 (GER14) reporter at the Rosa26 locus were mated. Females were super-ovulated and zygotes harvested. Zyotes were electroporated with various CRISPR/Cas9 combinations (or Cre) and were cultured 96 hours to blastocyst stage. Blastocysts were imaged for Katushka2S (RFP) fluorescence and scored for fluorescence signal. Blastocysts were then PCR amplified for the reporter allele and Sanger sequenced. ICE analysis tool from Synthego was used to score edits. NOTE: Some blasts did not PCR amplify well and it could not be determined if they had edits. Total blasts analyzed expressing the fluorescent reporter, 46-164. Total blast analyzed by Sanger sequencing, 39-142.

Peptide Shuttle optimization to deliver Cas9 RNP to human airway epithelia cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
Delivery of Cas9 RNP targeting human CFTR in Primary human epithelia cells. Gene editing efficiency was determined by percentage of NGS reads that showed an indel

Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
McCray Paul B  Last Updated Date: 2020-11-02
 
In vivo shuttle peptide delivery of Cas9 and Cas12a RNPs in mouse airway epithelia. Gene editing was quantified by the GFP+ cells in large and small airways following 1 delivery of GFP protein by GFP positive cells compared to DAPI stained cells.

Testing AAV5 for activation of tdTomato in HEK293T cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Gao Guang-Ping  Last Updated Date: 2020-10-20
 
AAV shuttle plasmids expressing SpCas9 and guide RNAs targeting the Ai9 transgene were tested in HEK293T cells by transient transfection. Both delivery and gene editing were detected by fluorescence.

FSD114

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD115

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD116d1

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168d11

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168d12

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD188

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD189

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD191

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD196

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD199

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD215

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD235

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD286

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD293

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD308

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD347

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD360

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD362

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57d1

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX1

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX6

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

AAV.pU1a-SpCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS

g-45_C12a

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
gRNA targeting CFTR inton-22-23

McCray_CFTR_Cas9_Guide1

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Targets endogenous human locus

sgAi9R

Guide  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
This sgRNA targets the Ai9 and related transgenes

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
GFP (D5.1) XP Rabbit mAb antibody

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
Anti-RFP (Rabbit) Polyclonal Antibody

Heaney-SATC_McCray-Validation_Lung Inflation and Fixation Protocol

Protocol  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
Procedure for mouse lung inflation and fixation.

FS48

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD121

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD132

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD160

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD240

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD259

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD260

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD283

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD284

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD287

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD289

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD297

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD303

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD304

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD305

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD307

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD310

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD316

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD334

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD339

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD365

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD368

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD94

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD97

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX7

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

S10D

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

S10-MOD

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

pH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)

Vector  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP

g-loxP2_C9

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
This sgRNA targets the Ai9 and related transgenes

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
Anti-RFP (RABBIT) Antibody

Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were injected into the intra-striatum of mTmG reporter mice and activation of GFP expression was imaged.

Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were injected into the striatum of TLR-MCV mice and the distribution of RNP was imaged.

STS120 (TLR1; Sp_t2:Sp_c20_TLR1)

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets traffic light reporter transgene

STS96 (PCSK9a; Sp_t2:Sp_c20_PCSK9a)

Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets endogenous mouse locus

HEK-293T-disrupted_GFP-mcherry-Puro

Model System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
HEK293T cells with an integrated reporter for TLR1 reporter editing. HEK293T is an epithelial-like cell that was isolated from the kidney of a patient.

Hepa1-6

Model System  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Stable mouse cell line of liver epithelial cells.

RRID:AB_1196615 

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Other Id: AB_1196615 RRID:AB_1196615
GFP (D5.1) XP Rabbit mAb antibody, Cell Signaling Technology

AB_2532994 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: AB_2532994 13-0300Ā 
Thermo-Fisher, Rat anti-GFAP

AB_141637 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: AB_141637 A21207
Invitrogen, Donkey anti-rabbit alexa fluor 594

AB_2813835 

Antibody  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to
Other Id: ab150155 AB_2813835
Abcam, Donkey anti-rat alexa fluour 647

Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-10-01
 
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain

Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were injected into the intra-striatum of mTmG reporter mice and activation of GFP expression was imaged.

Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain

Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were injected into the intra-striatum of mTmG reporter mice and activation of GFP expression was imaged.

Testing newly chemically modified crRNA and tracrRNA to activate the TLR1 reporter in human cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were delivered by electroporation to transgenic human HEK-293T cells harboring the TLR1 reporter. Gene editing was determined by reporter activation.

SpyCas9-3xNLS

Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
SpyCas9-3xNLS is type II-A Cas9 from Streptococcus pyogenes strain SF370. It was expressed from pMCSG7 bacterial expressing vector and purified from Escherichia coli Rosetta DE3 strain. SpyCas9 fused to 3 NLS: C-Myc-like NLS at the N-terminal SV40 NLS and Nucleoplasmin NLS at the C-terminal
Show Experiments (5)

Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain

STS118 (TLR-MCV1a; Sp_t2-PS:Sp_c20_TLR_MCV1a)

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets traffic light reporter transgene, CY3 labeled

STS204 (DNMT1; Sp_t2:Sp_c20_Dnmt1)

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
For endogenous locus

Mouse Embryonic Fibroblasts

Model System  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Primary cell line

RRID:AB_2536526 

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Other Id: RRID:AB_2536526 AB_2536526
GFP Recombinant Rabbit Monoclonal Antibody, Thermo Fisher Scientific #G10362

CasĪ¦-2

Genome Editor  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Compact editor of the Cas12j family identified in biggie phage from metagenomic assemblies

Cas12j-GFP10 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP16 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP23 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP2 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP7 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP7 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP8 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP12 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP14 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP1 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP3 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP4 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP6 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

HEK-293-TgEF1a-eGFP-BSD

Model System  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
HEK293 cells with lentiviral insertion of EF1a promoter driving expression of eGFP and SV40 promoter driving expression of BSD. HEK293 is an epithelial-like cell that was isolated from the kidney of a patient.

306-O12B blank

Delivery System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse]
Matched Fields: experimentName : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9
Combinatorial library cationic lipid nanoparticles

AsCas12a (Feldan Therapeutics)

Genome Editor  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides

AsCas12a (IDT and Feldan Therapeutics)

Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides

CM18-PTD4

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide

FS66d6

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD118

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168d20

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide

FSD168 Disul.

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD168 Scr.

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD186

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD190

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD227

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD228

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD234

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD317

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD318

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD322

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD333

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD335

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD359

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD361

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD364

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD366

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57d3

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57d4

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD57d5

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD63

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX5

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSX8

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

S85

Delivery System  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide

H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP

g-11_C9

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
sgRNA targeting CFTR exon 11

g-38330_C12a

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
gRNA targeting HPRT locus

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
Other Id:
Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000

Testing different ratios of Lipofectamine-RNPs after 24 hours for determination of positive control

Experiment  - [In Vitro] [Collaborative Opportunity Fund] [Human]
Matched Fields: description : Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles provided
Wilson Ross C. , Kiani Samira , Gong Shaoqin (Sarah)  Last Updated Date: 2023-12-18
 
Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles provided from the Gong Lab at the University of Wisconsin-Madison. The Gong lab conducted free radical polymerization of the monomer coating with (PEG)-acrylate to ensure that the RNP-NC be stable and able to conjugate different ligands. Two liver-targeted ligands were provided from the Gong Lab, RNP-NC attached to tri(GalNAc) and RNP-NC containing cell penetrating peptide (TAT). The tri(GalNAc) is known to enhance RNP-NC target to hepatocytes, whereas TAT will enhance target and uptake of RNP-NC in all liver cells such as Kupffer cells. These ligands are tagged with Atto-550 a fluorescent protein reporter for easier detection. The goal was to investigate cellular uptake of Cas9-gRNA nanocapsules using imaging after 24 hours and to determine the appropriate ratio for Lipofectamine-RNP positive control.

AAV9-CMV-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
AAV serotype 9 delivering CMV driven SaCas9

FSD115d1

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD195

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD197

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD262

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD95

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

AAV9-Ai9-sgRNA1 + sgRNA2

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus

AAVcc47-Ai9-sgRNA1 + sgRNA2

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus

AAVcc47-CMV-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
AAVcc47 delivering CMV driven SaCas9

FSD301

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

AAVcc47-Ai9-sgRNA1-CB-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus

Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were delivered by electroporation in presence of amphiphilic peptide to transgenic human HEK-293T cells harboring the TLR-MCV1 reporter. Gene editing was determined by reporter activation.

Testing newly chemically modified crRNA and tracrRNA in mouse Neuro 2A cells

Experiment  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Sontheimer Erik J  Last Updated Date: 2021-04-15
 
Chemically modified crRNA and tracrRNA were delivered by electroporation to mouse Neuro2A cells. Editing activity was determined by Sanger sequencing

SpyCas9-3xNLS

Genome Editor  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
SpyCas9-3xNLS is type II-A Cas9 from Streptococcus pyogenes strain SF370. It was expressed from pMCSG7 bacterial expressing vector and purified from Escherichia coli Rosetta DE3 strain. SpyCas9 fused to 3 NLS: C-Myc-like NLS at the N-terminal SV40 NLS and Nucleoplasmin NLS at the C-terminal

STS118 (TLR-MCV1a; Sp_t2:Sp_c20_TLR_MCV1a)

Guide  - [In Vitro] [Delivery Systems Initiative] [Human]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets traffic light reporter transgene

STS135 (PCSK9c; Sp_t2:Sp_c20_PCSK9c)

Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Targets endogenous mouse locus

STS159 (mTmG; Sp_t2-PS:Sp_c0_mTmG)

Guide  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
This sgRNA targets the mTmG transgene

Neuro 2A

Model System  - [In Vitro] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Neuro-2a cells are mouse neuroblasts with neuronal and amoeboid stem cell morphology isolated from brain tissue.

TLR-MCV1 mouse

Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
TLR-MCV1 transgene knocked into Rosa26 locus

RRID:AB_2798820 

Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors
Other Id: RRID:AB_2798820 AB_2798820
Cas9 (S. pyogenes) (E7M1H) XPĀ® Rabbit mAb antibody, Cell Signaling Technology

Testing newly discovered Biggie Phage editors in human cells

Experiment  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Banfield Jillian , Doudna Jennifer A  Last Updated Date: 2020-10-16
 

CasĪ¦-3

Genome Editor  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Compact editor of the Cas12j family identified in biggie phage from metagenomic assemblies

Cas12j-GFP19 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP22 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP3 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP5 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP9 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

AAV9-Ai9-sgRNA2-CB-SaCas9

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus

CasĪ¦-1

Genome Editor  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Compact editor of the Cas12j family identified in biggie phage from metagenomic assemblies

pPP441

Vector  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Plasmid containing Homo sapiens codon optimized CasΦ-2 and spacer for editing in human cells.

Cas12j-GFP1 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP21 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP2 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP4 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP5 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP5 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP8 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

SapI-GG stuffer (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Non-targeting guide RNA targeting compatible with CasĪ¦-1

SapI-GG stuffer (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Non-targeting guide RNA targeting compatible with CasĪ¦-2

SapI-GG stuffer (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Non-targeting guide RNA targeting compatible with CasĪ¦-3

FSD237

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD238

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD315

Delivery System  - [In Vivo, In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia
Show Experiments (7)

Peptide Shuttle optimization to deliver Cas9 RNP to human airway epithelia cells
Peptide Shuttle optimization to deliver Cas12a RNP to human airway epithelia cells
Delivery of Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting CFTR locus
Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to human airway epithelial cells cultured at the air liquid interface targeting B2M locus
Delivery of ABE8e-Cas9 RNP using Shuttle peptide candidates to rhesus monkey airway epithelial cells cultured at the air liquid interface targeting CCR5 locus.
Inhibitory effect of Cas9 RNP alone on S10- or FSDS315-mediated GFP protein delivery to HeLa cells
Amphiphilic Peptides Deliver Base Editor RNPs to Rhesus Monkey Airway

RNP-NC-no ligand

Delivery System  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Mouse]
Matched Fields: study : Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain experimentName : Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.
The nanocapsule is a thin glutathione (GSH)-cleavable covalently crosslinked polymer coating around a preassembled ribonucleoprotein (RNP) complex between a Cas9 nuclease and an sgRNA.

AAVcc47_pTR_self comp 2xU6-Ai9 guides

Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene

pPP394

Vector  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Plasmid containing Homo sapiens codon optimized CasΦ-1 and spacer for editing in human cells.

pPP444

Vector  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Plasmid containing Homo sapiens codon optimized CasΦ-3 and spacer for editing in human cells.

Cas12j-GFP10 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP11 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP13 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP14 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

Cas12j-GFP15 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP17 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP18 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP20 (CasĪ¦-1)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-1

Cas12j-GFP6 (CasĪ¦-2)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-2

Cas12j-GFP9 (CasĪ¦-3)

Guide  - [In Vitro] [Genome Editors] [Human]
Matched Fields: study : Expanding CRISPR-Cas Editing Technology through Exploration of Novel Cas Proteins and DNA Repair Systems
Guide targeting eGFP compatible with CasĪ¦-3

FSD114d1

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD285

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD321

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

FSD63D1

Delivery System  - [In Vitro] [Collaborative Opportunity Fund, Delivery Systems Initiative] [Human, Rhesus macaque]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Shuttle peptide used to deliver reagents to airway epithelia

Imaging quantification of transfection efficiency with varying dosages of nanoparticles encapsulated with Cas9/sgRNA RNP on the liver-on-chip model system

Experiment  - [In Vitro] [Collaborative Opportunity Fund] [Human]
Matched Fields: description : Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles.
Wilson Ross C. , Kiani Samira , Gong Shaoqin (Sarah)  Last Updated Date: 2023-12-18
 
Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles. Two liver-targeted ligands were provided from the Gong Lab, RNP-NC attached to tri(GalNAc) and RNP-NC containing cell penetrating peptide (TAT). The triGalNAc is known to enhance RNP-NC target to hepatocytes, whereas TAT will enhance target and uptake of RNP-NC in all liver cells such as Kupffer cells. These ligands are tagged with Atto-550 a fluorescent protein reporter for easier detection. Liver microtissue with a monoculture of primary human hepatocytes (PHH) was used. Each well of the liver-on-a-chip typically is seeded with 6.0 Ɨ 10^5 hepatocytes 16 hours prior to the addition of RNP-NCs with flow of 1.0 Āµl/s through the liver 3D microtissues. Lipofectamine ā€“ RNP complex was prepared using Lipofectamine 2000 Transfection Reagent with 1:1 weight to weight ratio after optimization of transfection efficiency. The goal of the experiment was to examine transfection efficiency by testing two doses 2.4 ug and 24 ug RNP-NCs [tri(GalNAc), TAT] with the help of imaging. Transfection of 24Āµg RNP-NC shows higher uptake when compared to2.4Āµg RNP-NC.

mTmG mouse (congenic)

Model System  - [In Vivo] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: study : Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors experimentName : Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia
mTmG is a double-fluorescent reporter transgenic mouse which expresses membrane-targeted tdTomato flanked by loxP sequences, followed by membrane-targeted GFP. After genomic cleavage by Cas9 at two sites, or Cre recombinase between loxP sites, tdTomato expression is lost and GFP is expressed.
Show Experiments (6)

Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center

306-O12B

Delivery System  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: experimentName : Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear
Combinatorial library cationic lipid nanoparticles

SpCas9

Genome Editor  - [In Vivo, In Vitro] [Delivery Systems Initiative, Biological Systems] [Human, Mouse]
Matched Fields: study : Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases
HA-SV40NLS-SpCas9-SV40NLS

Quantification of transfection efficiency by flow cytometry with varying dosages of nanoparticles encapsulated with Cas9/sgRNA RNP on liver-on-chip model system

Experiment  - [In Vitro] [Collaborative Opportunity Fund] [Human]
Matched Fields: description : Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles.
Wilson Ross C. , Kiani Samira , Gong Shaoqin (Sarah)  Last Updated Date: 2023-12-18
 
Liver-on-a-chip was used to examine the cellular uptake of CRISPR/Cas9 encapsulated nanoparticles. Two liver-targeted ligands were provided from the Gong Lab, RNP-NC attached to tri(GalNAc) and RNP-NC containing cell penetrating peptide (TAT). The triGalNAc is known to enhance RNP-NC target to hepatocytes, whereas TAT will enhance target and uptake of RNP- NC in all liver cells such as Kupffer cells. These ligands are tagged with Atto-550 a fluorescent protein reporter for easier detection. Liver microtissue with a monoculture of primary human hepatocytes (PHH) was used. Each well of the liver-on-a-chip typically is seeded with 6.0 Ɨ 10^5 hepatocytes 16 hours prior to the addition of RNP-NCs with flow of 1.0 Āµl/s through the liver 3D microtissues. Lipofectamine ā€“ RNP complex was prepared using Lipofectamine 2000 Transfection Reagent with 1:1 weight to weight ratio after optimization of transfection efficiency. The goal of the experiment was to quantify transfection efficiency by testing two doses 2.4 ug and 24 ug RNP-NCs [tri(GalNAc), TAT] using flow cytometry. Transfection of 24Āµg RNP-NC shows higher uptake when compared to2.4Āµg RNP-NC.

Alt-RĀ® S.p. Cas9 Nuclease V3

Genome Editor  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
Matched Fields: study : Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides
Recombinant S. pyogenes Cas9 nuclease, purified from an E. coli strain expressing the nuclease. Contains nuclear localization sequence (NLS) and C-terminal 6-His tag. Provided in solution at 10 Āµg/ĀµL. 100 Āµg of Cas9 nuclease = 610 pmol.
Show Experiments (9)

Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia
Validating Traffic Light Reporter 2 (TLR2) Mouse Model in Heterozygous Blastocysts using SpCas9
Validating Traffic Light Reporter 2 (TLR2) Mouse Model in Homozygous Blastocysts using SpCas9
Validating Gene Editing Reporter 12 (GER12) Mouse Model in Heterozygous Blastocysts
Validating Traffic Light Reporter 7 (TLR7) Mouse Model for NHEJ in Heterozygous Blastocysts
Validating Gene Editing Reporter 14 (GER14) Mouse Model in Heterozygous Blastocysts
Validating Gene Editing Reporter 18 (GER18) Mouse Model in Heterozygous Blastocysts
Validating Gene Editing Reporter 19 (GER19) Mouse Model in Heterozygous Blastocysts
Validating Traffic Light Reporter 7 (TLR7) Mouse Model for HDR in Heterozygous Blastocysts

320 results for crispr

Category Name Description Source View Associated..
Publication CRISPR-CasĪ¦ from huge phages is a hypercompact genome editor. CRISPR-Cas systems are found widely in prokaryotes, where they provide adaptive immunity against virus infection and plasmid transformation. We describe a minimal functional CRISPR-Cas system, comprising a single ~70-kilodalton protein, CasĪ¦, and a CRISPR array, encoded exclusively in the genomes of huge bacteriophages. CasĪ¦ uses a single active site for both CRISPR RNA (crRNA) processing and crRNA-guided DNA cutting to target foreign nucleic acids. This hypercompact system is active in vitro and in human and plant cells with expanded target recognition capabilities relative to other CRISPR-Cas proteins. Useful for genome editing and DNA detection but with a molecular weight half that of Cas9 and Cas12a genome-editing enzymes, CasĪ¦ offers advantages for cellular delivery that expand the genome editing toolbox.
Project Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides The proposed research is relevant to the public health because genetic and acquired diseases affecting the airways pose major disease and economic burdens. By advancing the delivery of gene editing tools, it may be possible to therapeutically modify the cells lining the airways. This novel strategy has implications for the treatment of both monogenetic and acquired lungs disease, and may have applications for other somatic cell therapies.
Project Enabling Nanoplatforms for Targeted In Vivo Delivery of CRISPR/Cas9 Riboncleoproteins in the Brain In vivo genome editing using CRISPR/Cas9 is anticipated to be the next wave of therapeutics for various major health threats, including neurodegenerative diseases. However, to date, very few Cas9-gRNA ribonucleoprotein in vivo delivery methods have been reported, and delivery to the brain has been particularly challenging. The unique nanocapsules we plan to develop will ultimately enable high efficiency neuron-targeted genome editing in the brain, thereby offering new hope to treat devastating neurodegenerative diseases.
Experiment Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain. Nanocapusules carrying CRISPR Cas9 RNP with guide RNA targeting the stop sequence in the Ai14 transgene are intracerebrally delivered to Ai14 mice and gene editing is measured by gain of tdTomato protein expression.
Project Develop Combinatorial Non-Viral and Viral CRISPR Delivery for Lung Diseases Efficacy and safety limitations in current gene editing technologies have hindered efforts to treat genetic lung diseases. This proposal seeks to develop and validate a combinatorial delivery approach that uses non-viral and viral vehicles to efficiently transport gene editing tools to disease-relevant cells in the lung. Completion of our work will establish safe and effective delivery vehicles that will guide the design of future gene therapies for genetic disorders.
Publication A bacterial cytidine deaminase toxin enables CRISPR-free mitochondrial base editing. Bacterial toxins represent a vast reservoir of biochemical diversity that can be repurposed for biomedical applications. Such proteins include a group of predicted interbacterial toxins of the deaminase superfamily, members of which have found application in gene-editing techniques1,2. Because previously described cytidine deaminases operate on single-stranded nucleic acids3, their use in base editing requires the unwinding of double-stranded DNA (dsDNA)-for example by a CRISPR-Cas9 system. Base editing within mitochondrial DNA (mtDNA), however, has thus far been hindered by challenges associated with the delivery of guide RNA into the mitochondria4. As a consequence, manipulation of mtDNA to date has been limited to the targeted destruction of the mitochondrialĀ genome by designer nucleases9,10.Here we describe an interbacterial toxin, which we name DddA, that catalyses the deamination of cytidines within dsDNA. We engineered split-DddA halves that are non-toxic and inactive until brought together on target DNA by adjacently bound programmable DNA-binding proteins. Fusions of the split-DddA halves, transcription activator-like effector array proteins, and a uracil glycosylase inhibitor resulted in RNA-free DddA-derived cytosine base editors (DdCBEs) that catalyse Cā€¢G-to-Tā€¢A conversions in human mtDNA with high target specificity and product purity. We used DdCBEs to model a disease-associated mtDNA mutation in human cells, resulting in changes in respiration rates and oxidative phosphorylation. CRISPR-free DdCBEs enable the precise manipulation of mtDNA, rather than the elimination of mtDNA copies that results from its cleavage by targeted nucleases, with broad implications for the study and potential treatment of mitochondrial disorders.
Publication CHANGE-seq reveals genetic and epigenetic effects on CRISPR-Cas9 genome-wide activity. Current methods can illuminate the genome-wide activity of CRISPR-Cas9 nucleases, but are not easily scalable to the throughput needed to fully understand the principles that govern Cas9 specificity. Here we describe 'circularization for high-throughput analysis of nuclease genome-wide effects by sequencing' (CHANGE-seq), a scalable, automatable tagmentation-based method for measuring the genome-wide activity of Cas9 in vitro. We applied CHANGE-seq to 110 single guide RNA targets across 13 therapeutically relevant loci in human primary T cells and identified 201,934 off-target sites, enabling the training of a machine learning model to predict off-target activity. Comparing matched genome-wide off-target, chromatin modification and accessibility, and transcriptional data, we found that cellular off-target activity was two to four times more likely to occur near active promoters, enhancers and transcribed regions. Finally, CHANGE-seq analysis of six targets across eight individual genomes revealed that human single-nucleotide variation had significant effects on activity at ~15.2% of off-target sites analyzed. CHANGE-seq is a simplified, sensitive and scalable approach to understanding the specificity of genome editors.
Publication Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia. The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSAmT/mG mice. We observe no evidence of short-term toxicity with a widespread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells.
Project Enhancing CRISPR Gene Editing in Somatic Tissues by Chemical Modification of Guides and Donors RNA-guided CRISPR genome editing systems promise to revolutionize the treatment of inherited disease. Safe, effective, and target-tissue-specific delivery of the guide RNA that directs editing is a critical hurdle in the development of clinical applications for engineered CRISPR systems. Using strategies validated for the delivery of other categories of nucleic acid therapeutics, we have established a framework for complete chemical modification of CRISPR guides, thereby conferring in vivo stability and effective biodistribution properties. The proposed research will optimize these guides, as well as other editing components, for clinical use.
Show Experiments (11)

Testing newly chemically modified crRNA and tracrRNA to activate the TLR reporter in human cells
Testing newly chemically modified crRNA and tracrRNA in mTmG mouse embryonic fibroblasts
Testing newly chemically modified crRNA and tracrRNA to activate the TLR1 reporter in human cells
Testing newly chemically modified crRNA and tracrRNA in mouse Hepa 1-6 cells
Testing newly chemically modified crRNA and tracrRNA in mouse Neuro 2A cells
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA using the S10 shuttle peptide to activate the mTmG reporter in mouse brain
Delivery of unmodified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, extended PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Delivery of RNP containing chemically modified crRNA C20 with chemically modified tracrRNA T2-PS to determine the RNP distribution in TLR-MCV mouse brain
Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing Center

Publication Self-delivering, chemically modified CRISPR RNAs for AAV co-delivery and genome editing in vivo. Guide RNAs offer programmability for CRISPR-Cas9 genome editing but also add challenges for delivery. Chemical modification, which has been key to the success of oligonucleotide therapeutics, can enhance the stability, distribution, cellular uptake, and safety of nucleic acids. Previously, we engineered heavily and fully modified SpyCas9 crRNA and tracrRNA, which showed enhanced stability and retained activity when delivered to cultured cells in the form of the ribonucleoprotein complex. In this study, we report that a short, fully stabilized oligonucleotide (a 'protecting oligo'), which can be displaced by tracrRNA annealing, can significantly enhance the potency and stability of a heavily modified crRNA. Furthermore, protecting oligos allow various bioconjugates to be appended, thereby improving cellular uptake and biodistribution of crRNA in vivo. Finally, we achieved in vivo genome editing in adult mouse liver and central nervous system via co-delivery of unformulated, chemically modified crRNAs with protecting oligos and AAV vectors that express tracrRNA and either SpyCas9 or a base editor derivative. Our proof-of-concept establishment of AAV/crRNA co-delivery offers a route towards transient editing activity, target multiplexing, guide redosing, and vector inactivation.
Experiment [Validation] Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear Delivery of CRISPR/Cas9 via bioreducible l