93 results for Epithelium

Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia.   - Experiment - [In Vivo]
Matched on: description ->  Delivery of GFP via shuttle peptides to mouse airway epithelium via nasal instilation. generatedDescription ->  Delivery of GFP via shuttle peptides to mouse airway epithelium via nasal instilation. tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
McCray Paul B
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.
View Associated: 
BALB/c mouse   - Model System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
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).
View Associated: 
Alt-R® S.p. Cas9 Nuclease V3   - Genome Editor
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
View Associated: 
GFP-NLS   - Genome Editor
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
Nuclear targeted GFP
View Associated: 
Testing AAV5 for activation of tdTomato in mouse airway   - Experiment - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
Gao Guang-Ping
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
View Associated: 
Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia   - Experiment - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
McCray Paul B
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.
View Associated: 
AsCas12a (IDT and Feldan Therapeutics)   - Genome Editor
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
View Associated: 
g-loxP2_C9   - Guide - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
This sgRNA targets the Ai9 and related transgenes
View Associated: 
SpCas9   - Genome Editor
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
HA-SV40NLS-SpCas9-SV40NLS
View Associated: 
AAV.pU1a-SpCas9   - Vector
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS
View Associated: 
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)   - Vector
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP
View Associated: 
g-loxPbot_C12a   - Guide - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
This sgRNA targets the Ai9 and related transgenes at two sites
View Associated: 
mTmG mouse   - Model System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Lung epitheliumEpithelium of mucosa ; Epithelium ; Foregut epithelium ; Endo-epithelium
ROSAmT/mG 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
View Associated: 
S10   - Delivery System - [In Vivo, In Vitro]
Matched on: tissueTerm ->  Epithelium of bronchiole ; Epithelium of main bronchus Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
Shuttle peptide used to deliver reagents to airway epithelia
View Associated: 
AB_1196615    - Antibody - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Columnar epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
GFP (D5.1) XP Rabbit mAb antibody
View Associated: 
AB_2209751    - Antibody - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Ecto-epitheliumEpithelium ; Endo-epithelium ; Lung epithelium
Anti-RFP (RABBIT) Antibody
View Associated: 
Ai9 mouse   - Model System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Ecto-epithelium ; Lung epitheliumEpithelium ; Endo-epithelium
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.
View Associated: 
mTmG mouse (congenic)   - Model System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Ecto-epithelium ; Lung epitheliumEpithelium ; Endo-epithelium
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.
View Associated: 
SpCas9   - Genome Editor
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Lung epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
View Associated: 
Ai9 mouse (BCM)   - Model System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Ecto-epithelium ; Lung epitheliumEpithelium ; Endo-epithelium
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.
View Associated: 
[] Validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides   - Experiment - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Lung epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
Heaney Jason D
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
View Associated: 
D10   - Delivery System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Lung epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
View Associated: 
D237   - Delivery System - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Lung epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
View Associated: 
SpyCas9 g-loxP2_C9   - Guide - [In Vivo]
Matched on: tissueTerm ->  Epithelium of main bronchus ; Epithelium of bronchiole Tissue/CellType Term Synonyms ->  Epithelium of mucosa ; Lung epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
This sgRNA targets the mTmG transgene
View Associated: 
AAVcc47_pTR_self comp 2xU6-Ai9 guides   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
View Associated: 
Cre Recombinase dose escalation study in Ai9 mice   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
Asokan Aravind
A single stranded cmv cre cassette was packaged into AAV9 or AAVcc47 and injected intravenously in Ai9 mice. We injected n=3 at three different doses (1e10, 1e11, 1e12 vg) and harvested organs 4 weeks post injection. Fluorescence intensity in liver, heart, and skeletal muscle was quantified with tiff based images in Image J and neuronal transduction from each vector was quantified at the 1e12vg dose by counting the number of tdTomato+ neurons and number of NeuN+ cells from multiple sections and images.
View Associated: 
306-O12B   - Delivery System - [In Vivo, In Vitro]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Unilaminar epithelium
Combinatorial library cationic lipid nanoparticles
View Associated: 
Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
Asokan Aravind
C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding an mCherry reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
View Associated: 
AAVcc84-GFP   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter
View Associated: 
[] Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epithelium ; Foregut epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
Heaney Jason D
Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery. Detection of editing in non-target tissues.
View Associated: 
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]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
Asokan Aravind
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.
View Associated: 
AAV9-Ai9-sgRNA1-CB-SaCas9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
View Associated: 
AAV9-Ai9-sgRNA2-CB-SaCas9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
View Associated: 
AAVcc47-Ai9-sgRNA1-CB-SaCas9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
View Associated: 
AAVcc47-Ai9-sgRNA1 + sgRNA2   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
View Associated: 
Testing preparation for validation at The Jackson Laboratory Small Animal Testing Center   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Sontheimer Erik J
Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
View Associated: 
Cre recombinase   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
Cre recombinase delivered by AAVcc47-Cre viral vector
View Associated: 
AAVcc47-CMV-Cre   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAVcc47 delivering CMV Cre Recombinase
View Associated: 
RNP-NC-CPP   - Delivery System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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
View Associated: 
CleanCap® Fluc   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Digestive tract epithelium ; Columnar epitheliumEpithelium ; Unilaminar epithelium ; Endo-epithelium
Firefly luciferase mRNA capped using CleanCap. It is polyadenylated, modified with 5-methoxyuridine and optimized for mammalian systems. It mimics a fully processed mature mRNA.
View Associated: 
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]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
Asokan Aravind
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.
View Associated: 
AAV9_pTR_self comp 2xU6-Ai9 guides   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
View Associated: 
Testing gRNA sequence and gRNA scaffold modified in Ai9 mice.   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Deverman Benjamin E
3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2 were codelivered intravenously to adult male and female Ai9 mice. Editing was assessed in brain sections 4 weeks later.
View Associated: 
AAVcc47-Cre   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV2/5 expressing Cre recombinase
View Associated: 
AAV9-CMV-Cre   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV serotype 9 delivering CMV Cre Recombinase
View Associated: 
Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Chen Zheng-Yi
Delivery of Cas9/sgRNA mRNA via new LNPs to the cochlea by cochleostomy and gene editing is measured by percentage of tdTomato positive cells.
View Associated: 
AAV9-GFP   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 self complementary vector expressing GFP driven by CBh promoter
View Associated: 
AAVcc81-GFP   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter
View Associated: 
SauCas9   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Columnar epithelium ; Foregut epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
View Associated: 
CleanCap® Cas9   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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.
View Associated: 
sg298   - Guide - [In Vivo, In Vitro]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
This sgRNA targets the Ai9 and related transgenes at multiple sites
View Associated: 
AB_10015251    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Anti-Myosin VIIa antibody, Proteus Biosciences
View Associated: 
AB_1196615    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
GFP (D5.1) XP Rabbit mAb antibody, Cell Signaling Technology
View Associated: 
[] Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Heaney Jason D
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
View Associated: 
Ai14 mouse   - Model System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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.
View Associated: 
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]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
Asokan Aravind
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.
View Associated: 
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]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
Asokan Aravind
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.
View Associated: 
AAV9-Ai9-sgRNA1 + sgRNA2   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
View Associated: 
AAVcc47-Ai9-sgRNA2-CB-SaCas9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus
View Associated: 
AAVcc47-CMV-SaCas9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Unilaminar epithelium
AAVcc47 delivering CMV driven SaCas9
View Associated: 
AB_2286684    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology
View Associated: 
CleanCap® Cas9 mRNA (5moU)   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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.
View Associated: 
C57BL/6J mouse   - Model System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium ; Foregut epithelium
C57BL/6J WT mouse
View Associated: 
113-O12B   - Delivery System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Unilaminar epithelium
combinatorial library cationic lipid nanoparticles
View Associated: 
Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
Asokan Aravind
C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding a GFP reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
View Associated: 
C57BL/6 mouse (Asokan study)   - Model System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
View Associated: 
[] Second site validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Murray Stephen A
Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice
View Associated: 
AB_2532994    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Thermo-Fisher, Rat anti-GFAP
View Associated: 
AB_141637    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Invitrogen, Donkey anti-rabbit alexa fluor 594
View Associated: 
AB_141607    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Invitrogen, Donkey anti-mouse alexafluor 488
View Associated: 
306-O12B blank   - Delivery System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Combinatorial library cationic lipid nanoparticles
View Associated: 
BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds
View Associated: 
Testing new LNPs (lipid nanoparticles) for delivery of Fluc mRNA in adult mice   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Digestive tract epithelium ; Columnar epitheliumEpithelium ; Unilaminar epithelium ; Endo-epithelium
Chen Zheng-Yi
Delivery of firefly luciferase mRNA via new Lipid NanoParticles by tail vein injection into WT C57BL/6J mice targeting the Liver and delivery is measured by luciferase expression.
View Associated: 
AAV9-mCherry   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter
View Associated: 
AAVcc47-mCherry   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter
View Associated: 
306-S10   - Delivery System - [In Vivo, In Vitro]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epithelium ; Columnar epitheliumEpithelium ; Endo-epithelium ; Unilaminar epithelium
combinatorial library cationic lipid nanoparticles
View Associated: 
AAVcc47-SaCas9-Ai9   - Vector
Matched on: Tissue/CellType Term Synonyms ->  Columnar epithelium ; Foregut epitheliumEpithelium ; Endo-epithelium ; Meso-epithelium
AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter
View Associated: 
Ai14 mouse (congenic)   - Model System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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.
View Associated: 
SpyCas9-3xNLS   - Genome Editor
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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
View Associated: 
Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Gong Shaoqin (Sarah)
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.
View Associated: 
RNP-NC-RVG   - Delivery System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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
View Associated: 
AB_10711040    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Abcam, mouse anti-NeuN
View Associated: 
AB_2813835    - Antibody - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Abcam, Donkey anti-rat alexa fluour 647
View Associated: 
[] Second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Murray Stephen A
Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice
View Associated: 
[] Repeat experiment of second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear   - Experiment - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
Murray Stephen A
Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice
View Associated: 
RNP-NC-no ligand   - Delivery System - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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.
View Associated: 
sg298   - Guide - [In Vivo]
Matched on: Tissue/CellType Term Synonyms ->  Ecto-epitheliumEpithelium
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
View Associated: 

93 results for Epithelium

Category Type Name Description View Associated..
Experiment In Vivo Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia. 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.
Model System Animal BALB/c mouse 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).
Genome Editor Cas Nuclease Alt-R® S.p. Cas9 Nuclease V3
Genome Editor Reporter GFP-NLS Nuclear targeted GFP
Experiment In Vivo Testing AAV5 for activation of tdTomato in mouse airway 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
Experiment In Vivo Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia 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.
Genome Editor Cas Nuclease AsCas12a (IDT and Feldan Therapeutics)
Guide g-loxP2_C9 This sgRNA targets the Ai9 and related transgenes
Genome Editor Cas Nuclease SpCas9 HA-SV40NLS-SpCas9-SV40NLS
Vector Viral Vector AAV.pU1a-SpCas9 Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS
Vector Viral Vector H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1) AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP
Guide g-loxPbot_C12a This sgRNA targets the Ai9 and related transgenes at two sites
Model System Animal mTmG mouse ROSAmT/mG 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
Delivery System Amphiphilic peptide S10 Shuttle peptide used to deliver reagents to airway epithelia
Antibody AB_1196615  GFP (D5.1) XP Rabbit mAb antibody
Antibody AB_2209751  Anti-RFP (RABBIT) Antibody
Model System Animal Ai9 mouse 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.
Model System Animal mTmG mouse (congenic) 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.
Genome Editor Cas Nuclease SpCas9
Model System Animal Ai9 mouse (BCM) 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.
Experiment In Vivo Validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides 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
Delivery System Amphiphilic peptide D10
Delivery System Amphiphilic peptide D237
Guide SpyCas9 g-loxP2_C9 This sgRNA targets the mTmG transgene
Vector Viral Vector AAVcc47_pTR_self comp 2xU6-Ai9 guides AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
Experiment In Vivo Cre Recombinase dose escalation study in Ai9 mice A single stranded cmv cre cassette was packaged into AAV9 or AAVcc47 and injected intravenously in Ai9 mice. We injected n=3 at three different doses (1e10, 1e11, 1e12 vg) and harvested organs 4 weeks post injection. Fluorescence intensity in liver, heart, and skeletal muscle was quantified with tiff based images in Image J and neuronal transduction from each vector was quantified at the 1e12vg dose by counting the number of tdTomato+ neurons and number of NeuN+ cells from multiple sections and images.
Delivery System Nanoparticle 306-O12B Combinatorial library cationic lipid nanoparticles
Experiment In Vivo Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding an mCherry reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
Vector Viral Vector AAVcc84-GFP AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter
Experiment In Vivo Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing. Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery. Detection of editing in non-target tissues.
Experiment In Vivo Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter) 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.
Vector Viral Vector AAV9-Ai9-sgRNA1-CB-SaCas9 AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAV9-Ai9-sgRNA2-CB-SaCas9 AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAVcc47-Ai9-sgRNA1-CB-SaCas9 AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAVcc47-Ai9-sgRNA1 + sgRNA2 AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
Experiment In Vivo Testing preparation for validation at The Jackson Laboratory Small Animal Testing Center Delivery of chemically modified, phosphorothioate (PS)-stabilized crRNA with chemically modified, PS-stabilized tracrRNA to activate the mTmG reporter in mouse brain
Genome Editor Recombinase Cre recombinase Cre recombinase delivered by AAVcc47-Cre viral vector
Vector Viral Vector AAVcc47-CMV-Cre AAVcc47 delivering CMV Cre Recombinase
Delivery System Nanoparticle RNP-NC-CPP 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
Genome Editor Reporter CleanCap® Fluc Firefly luciferase mRNA capped using CleanCap. It is polyadenylated, modified with 5-methoxyuridine and optimized for mammalian systems. It mimics a fully processed mature mRNA.
Experiment In Vivo Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter) 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.
Genome Editor Cas Nuclease SaCas9-Lagor
Vector Viral Vector AAV9_pTR_self comp 2xU6-Ai9 guides AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
Experiment In Vivo Testing gRNA sequence and gRNA scaffold modified in Ai9 mice. 3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2 were codelivered intravenously to adult male and female Ai9 mice. Editing was assessed in brain sections 4 weeks later.
Vector Viral Vector AAVcc47-Cre AAV2/5 expressing Cre recombinase
Vector Viral Vector AAV9-CMV-Cre AAV serotype 9 delivering CMV Cre Recombinase
Experiment In Vivo Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea Delivery of Cas9/sgRNA mRNA via new LNPs to the cochlea by cochleostomy and gene editing is measured by percentage of tdTomato positive cells.
Vector Viral Vector AAV9-GFP AAV2/9 self complementary vector expressing GFP driven by CBh promoter
Vector Viral Vector AAVcc81-GFP AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter
Genome Editor Cas Nuclease SauCas9
Genome Editor Cas Nuclease CleanCap® Cas9 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.
Guide sg298 This sgRNA targets the Ai9 and related transgenes at multiple sites
Antibody AB_10015251  Anti-Myosin VIIa antibody, Proteus Biosciences
Antibody AB_1196615  GFP (D5.1) XP Rabbit mAb antibody, Cell Signaling Technology
Guide Ai14 gRNA This sgRNA targets the Ai9 and related transgenes at multiple sites
Experiment In Vivo Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain 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
Genome Editor Cas Nuclease SaCas9
Model System Animal Ai14 mouse 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.
Experiment In Vivo Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CB promoter) 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.
Experiment In Vivo 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. 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.
Vector Viral Vector AAV9-Ai9-sgRNA1 + sgRNA2 AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
Vector Viral Vector AAVcc47-Ai9-sgRNA2-CB-SaCas9 AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAVcc47-CMV-SaCas9 AAVcc47 delivering CMV driven SaCas9
Antibody AB_2286684  Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology
Genome Editor Cas Nuclease CleanCap® Cas9 mRNA (5moU) 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.
Genome Editor Cas Nuclease sNLS-SpCas9-sNLS SpCas9 with N- and C-terminal SV40 NLS
Model System Animal C57BL/6J mouse C57BL/6J WT mouse
Delivery System Nanoparticle 113-O12B combinatorial library cationic lipid nanoparticles
Experiment In Vivo Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding a GFP reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
Model System Animal C57BL/6 mouse (Asokan study)
Experiment In Vivo Second site validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice
Antibody AB_2532994  Thermo-Fisher, Rat anti-GFAP
Antibody AB_141637  Invitrogen, Donkey anti-rabbit alexa fluor 594
Antibody AB_141607  Invitrogen, Donkey anti-mouse alexafluor 488
Delivery System Nanoparticle 306-O12B blank Combinatorial library cationic lipid nanoparticles
Vector Viral Vector BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2 Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds
Experiment In Vivo Testing new LNPs (lipid nanoparticles) for delivery of Fluc mRNA in adult mice Delivery of firefly luciferase mRNA via new Lipid NanoParticles by tail vein injection into WT C57BL/6J mice targeting the Liver and delivery is measured by luciferase expression.
Vector Viral Vector AAV9-CMV-SaCas9 AAV serotype 9 delivering CMV driven SaCas9
Vector Viral Vector AAV9-mCherry AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter
Vector Viral Vector AAVcc47-mCherry AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter
Delivery System Nanoparticle 306-S10 combinatorial library cationic lipid nanoparticles
Vector Viral Vector AAVcc47-SaCas9-Ai9 AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter
Model System Animal Ai14 mouse (congenic) 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.
Genome Editor Cas Nuclease SpyCas9-3xNLS 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
Experiment In Vivo 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.
Delivery System Nanoparticle RNP-NC-RVG 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
Antibody AB_10711040  Abcam, mouse anti-NeuN
Antibody AB_2813835  Abcam, Donkey anti-rat alexa fluour 647
Experiment In Vivo Second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice
Experiment In Vivo Repeat experiment of second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice
Delivery System Nanoparticle RNP-NC-no ligand 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.
Guide sg298 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
Vector Viral Vector BI28:AAV-GFAP-SaCas9-WPRE3-pA Novel engineered AAV BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein (GFAP) promoter