125 results for Epithelium

Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia.   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Delivery of GFP via shuttle peptides to mouse airway epithelium via nasal instilation.  -->  Delivery of GFP via shuttle peptides to mouse airway epithelium via nasal instilation.  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium 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.
Alt-R® S.p. Cas9 Nuclease V3   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium of mucosa  Epithelium  Foregut epithelium  Endo-epithelium
g-loxP2_C9   - Guide [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium of mucosa  Epithelium  Foregut epithelium  Endo-epithelium
This sgRNA targets the Ai9 and related transgenes
BALB/c mouse   - Model System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium 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).
Matched on:  -->  Epithelium of bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Santa Cruz, SC-25555 
Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000
AsCas12a (IDT and Feldan Therapeutics)   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium of mucosa  Epithelium  Foregut epithelium  Endo-epithelium
Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epithelia   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium 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.
GFP-NLS   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium of mucosa  Epithelium  Foregut epithelium  Endo-epithelium
Nuclear targeted GFP
sgAi9L   - Guide 
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
This sgRNA targets the Ai9 and related transgenes
sgAi9R   - Guide 
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
This sgRNA targets the Ai9 and related transgenes
mTmG mouse   - Model System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium 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
Cre recombinase   - Genome Editor  [Small Animal Testing Center (SATC), Delivery Systems Initiative]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Epithelium  Endo-epithelium  Lung epithelium  Meso-epithelium
Cre recombinase delivered by AAV (see vector details)
ssAAV5-sgB.saCas9   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Columnar epithelium  Epithelium  Endo-epithelium  Lung epithelium
BCM_ssAAV5-Sa_sgB   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Columnar epithelium  Epithelium  Endo-epithelium  Lung epithelium
This gRNA targets the Ai9 and related transgenes
Matched on:  -->  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Rockland Cat# 600-401-379 
Anti-RFP (RABBIT) Antibody
Matched on:  -->  Epithelium of bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Thermo Fisher Scientific, MA5-15257 
Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300
Matched on:  -->  Epithelium of bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Rockland, 600- 401-379 
Anti-RFP (Rabbit) Polyclonal Antibody
Matched on:  -->  Epithelium of bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Santa Cruz, SC-23950 
Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200
SpCas9   - Genome Editor  [Biological Systems]
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
HA-SV40NLS-SpCas9-SV40NLS
AAV.pU1a-SpCas9   - Vector 
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)   - Vector 
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP
g-loxPbot_C12a   - Guide [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Lung epithelium  Epithelium of mucosa  Epithelium  Foregut epithelium  Endo-epithelium
This sgRNA targets the Ai9 and related transgenes at two sites
SaCas9   - Genome Editor  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Columnar epithelium  Epithelium  Endo-epithelium  Lung epithelium
S10   - Delivery System [Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Columnar epithelium  Epithelium  Endo-epithelium  Lung epithelium
Shuttle peptide used to deliver reagents to airway epithelia
scAAV5-Cre-GFP   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of bronchiole  Epithelium of main bronchus  -->  Epithelium of mucosa  Columnar epithelium  Epithelium  Endo-epithelium  Lung epithelium
Matched on:  -->  Epithelium of main bronchus  -->  Columnar epithelium  Epithelium  Foregut epithelium  Ciliated epithelium  Endo-epithelium
Other Id:   Cell Signaling Technology Cat# 2956 
GFP (D5.1) XP Rabbit mAb antibody
D10   - Delivery System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
Ai9 mouse   - Model System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of bronchus  Epithelium of main bronchus  -->  Epithelium of mucosa  Ecto-epithelium  Epithelium  Endo-epithelium  Meso-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.
D237   - Delivery System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
SpCas9   - Genome Editor  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
BCM_ssAAV5-Sp_sgB   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This sgRNA targets the Ai9 and related transgenes
SpyCas9 g-loxP2_C9   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This sgRNA targets the mTmG transgene
ssAAV5-sgA+sgB-U1A.GFP   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
Ai9 mouse (BCM)   - Model System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Ecto-epithelium  Lung epithelium  Epithelium  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.
Ai9-SauSpyCas9 mouse   - Model System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
Ai9-SauSpyCas9 is a version of Ai9 that has a single guide RNA sites on both sides of the STOP cassette. This is a Cre reporter allele that 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. Ai9 mice express robust tdTomato fluorescence following Cre-mediated recombination.
[Validation] Validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic Peptides   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  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
ssAAV5-spCas9   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
BCM_ssAAV5-Sp_sgA   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This sgRNA targets the Ai9 and related transgenes
mTmG mouse (congenic)   - Model System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Ecto-epithelium  Lung epithelium  Epithelium  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.
[Validation] Validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Epithelium of main bronchus  Epithelium of bronchiole  -->  Epithelium of mucosa  Lung epithelium  Epithelium  Endo-epithelium  Meso-epithelium
Heaney Jason D
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
AAV   - Delivery System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Digestive tract epithelium  Columnar epithelium  Epithelium  Unilaminar epithelium  Endo-epithelium
See vector details
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 [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Columnar epithelium  Epithelium  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.
AAV9-Ai9-sgRNA1 + sgRNA2   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
AAVcc47_pTR_self comp 2xU6-Ai9 guides   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
113-O12B   - Delivery System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Unilaminar epithelium
combinatorial library cationic lipid nanoparticles
Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  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.
[Validation] Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Columnar epithelium  Foregut epithelium  Epithelium  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.
SauCas9   - Genome Editor  [Small Animal Testing Center (SATC)]
Matched on:  -->  Columnar epithelium  Foregut epithelium  Epithelium  Endo-epithelium  Meso-epithelium
Ai9 SaCas9 Guide B   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Columnar epithelium  Foregut epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This gRNA targets the Ai9 and related transgenes
STS159 (mTmG; Sp_t2:Sp_c20_mTmG)   - Guide [Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Epithelium
Targets mTmG and related reporter transgenes
RNP-NC-RVG   - Delivery System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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
AB_141607    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: A21202 
Invitrogen, Donkey anti-mouse alexafluor 488
CleanCap® Cas9 mRNA (5moU)   - Genome Editor  [Small Animal Testing Center (SATC)]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
sNLS-SpCas9-sNLS   - Genome Editor  [Small Animal Testing Center (SATC), Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Epithelium
SpCas9 with N- and C-terminal SV40 NLS
RNP-NC-CPP   - Delivery System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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
L2-modified   - Guide [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Epithelium
This gRNA targets the Ai9 and related transgenes
BI28:AAV-GFAP-SaCas9-WPRE3-pA   - Vector  [Small Animal Testing Center (SATC), Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Epithelium
Novel engineered AAV BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein (GFAP) promoter
AB_2286684    - Antibody [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: sc-17320 
Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CB promoter)   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Columnar epithelium  Epithelium  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.
AAV9_pTR_self comp 2xU6-Ai9 guides   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
AAVcc47-Ai9-sgRNA1-CB-SaCas9   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
AAV9-CMV-Cre   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV serotype 9 delivering CMV Cre Recombinase
AAV9-Ai9-sgRNA2-CB-SaCas9   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
AAVcc47-CMV-SaCas9   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAVcc47 delivering CMV driven SaCas9
sgAi9L   - Guide 
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This sgRNA targets the Ai9 and related transgenes
SpyCas9-3xNLS   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Epithelium
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
C57BL/6J mouse   - Model System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Foregut epithelium
C57BL/6J WT mouse
AAVcc47-CMV-Cre   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAVcc47 delivering CMV Cre Recombinase
Ai14 mouse   - Model System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
306-O12B   - Delivery System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Unilaminar epithelium
Combinatorial library cationic lipid nanoparticles
Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  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.
AAV9-GFP   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 self complementary vector expressing GFP driven by CBh promoter
C57BL/6 mouse (Asokan study)   - Model System [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AB_2813835    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: ab150155 
Abcam, Donkey anti-rat alexa fluour 647
RRID:AB_10711040    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Abcam, ab104224 
Mouse monoclonal [1B7] to NeuN - Neuronal Marker
AAVcc47-SaCas9-Ai9   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Columnar epithelium  Foregut epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter
[Validation] Repeat experiment of second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Murray Stephen A
Delivery of CRISPR/Cas9 editor via bioreducible lipid nanoparticle to the inner ear in Ai14 mice
sg298   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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
[Validation] Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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
R2-modified   - Guide [Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Epithelium
This gRNA targets the Ai9 and related transgenes
AB_10015251    - Antibody [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: 25-6790 
Anti-Myosin VIIa antibody, Proteus Biosciences
RRID:AB_2762834    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: ThermoFisher #A32794 
Alexa fluor 555, Donkey anti-rabbit
RRID:AB_141607    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Thermofisher #A21202 
Alexa fluor 488, Donkey anti-mouse
RNP-NC-no ligand   - Delivery System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
Ai14 gRNA   - Guide [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
This sgRNA targets the Ai9 and related transgenes at multiple sites
RRID:AB_10563941    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Invitrogen, R10367 
Polyclonal rabbit anti-RFP antibody
Testing new LNPs (lipid nanoparticles) for delivery of Fluc mRNA in adult mice   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Digestive tract epithelium  Columnar epithelium  Epithelium  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.
Testing gRNA sequence and gRNA scaffold modified in Ai9 mice.   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2   - Vector  [Small Animal Testing Center (SATC), Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Epithelium
Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds
L1-modified   - Guide [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Epithelium
This gRNA targets the Ai9 and related transgenes
AAVcc47-Cre   - Vector  [Small Animal Testing Center (SATC)]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV2/5 expressing Cre recombinase
Cre Recombinase dose escalation study in Ai9 mice   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  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.
Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochlea   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
AAV9-mCherry   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter
AAVcc47-mCherry   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter
AAVcc84-GFP   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter
Testing preparation for validation at The Jackson Laboratory Small Animal Testing Center   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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
[Validation] Second site validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Murray Stephen A
Delivery of CRISPR/Cas9 via RNP-loaded nanocages to the brain in Ai14 mice
AB_2532994    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: 13-0300  
Thermo-Fisher, Rat anti-GFAP
AB_141637    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: A21207 
Invitrogen, Donkey anti-rabbit alexa fluor 594
RRID:AB_2633281    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Invitrogen Product #A32732 
Goat anti-rabbit IgG-Alexa Fluor 555
CleanCap® Fluc   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Digestive tract epithelium  Columnar epithelium  Epithelium  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.
306-O12B blank   - Delivery System [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Combinatorial library cationic lipid nanoparticles
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter)   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Columnar epithelium  Epithelium  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.
Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter)   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Columnar epithelium  Epithelium  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.
AAV9-Ai9-sgRNA1-CB-SaCas9   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
AAVcc47-Ai9-sgRNA1 + sgRNA2   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
AAVcc47-Ai9-sgRNA2-CB-SaCas9   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium  Unilaminar epithelium
AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus
AAVcc81-GFP   - Vector  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Meso-epithelium
AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter
306-S10   - Delivery System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Columnar epithelium  Epithelium  Endo-epithelium  Unilaminar epithelium
combinatorial library cationic lipid nanoparticles
Ai9 SaCas9 Guide A   - Guide [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Columnar epithelium  Foregut epithelium  Epithelium  Endo-epithelium  Meso-epithelium
This gRNA targets the Ai9 and related transgenes
CleanCap® Cas9   - Genome Editor  [Delivery Systems Initiative]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
STS204 (DNMT1; Sp_t2:Sp_c20_Dnmt1)   - Guide [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
For endogenous locus
Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.   - Experiment [Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.
AB_2209751    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Rockland Cat# 600-401-379 
Anti-RFP (RABBIT) Antibody
AB_10711040    - Antibody [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: ab104224  
Abcam, mouse anti-NeuN
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id:   GeneTex Product # GTX00837 
Chicken Polyclonal anti-NeuN antibody
RRID:AB_2762843    - Antibody 
Matched on:  -->  Ecto-epithelium  Epithelium
Other Id: Invitrogen Product # A32931 
Goat anti-chicken IgG-Alexa Fluor 488
[Validation] Second site validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear   - Experiment [Small Animal Testing Center (SATC)] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
Murray Stephen A
Delivery of CRISPR/Cas9 via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice
sg298   - Guide [Delivery Systems Initiative] - [In Vivo, In Vitro]
Matched on:  -->  Ecto-epithelium  Epithelium
This sgRNA targets the Ai9 and related transgenes at multiple sites
Ai14 mouse (congenic)   - Model System [Small Animal Testing Center (SATC), Delivery Systems Initiative] - [In Vivo]
Matched on:  -->  Ecto-epithelium  Epithelium
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.

125 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.
Genome Editor Cas Nuclease Alt-R® S.p. Cas9 Nuclease V3
Guide g-loxP2_C9 This sgRNA targets the Ai9 and related transgenes
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).
Antibody   Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000
Genome Editor Cas Nuclease AsCas12a (IDT and Feldan Therapeutics)
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 Reporter GFP-NLS Nuclear targeted GFP
Guide sgAi9L This sgRNA targets the Ai9 and related transgenes
Guide sgAi9R This sgRNA targets the Ai9 and related transgenes
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
Genome Editor Recombinase Cre recombinase Cre recombinase delivered by AAV (see vector details)
Vector Viral Vector ssAAV5-sgB.saCas9
Guide BCM_ssAAV5-Sa_sgB This gRNA targets the Ai9 and related transgenes
Antibody   Anti-RFP (RABBIT) Antibody
Antibody   Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300
Antibody   Anti-RFP (Rabbit) Polyclonal Antibody
Antibody   Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200
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
Genome Editor Cas Nuclease SaCas9
Delivery System Amphiphilic peptide S10 Shuttle peptide used to deliver reagents to airway epithelia
Vector Viral Vector scAAV5-Cre-GFP
Antibody   GFP (D5.1) XP Rabbit mAb antibody
Delivery System Amphiphilic peptide D10
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.
Delivery System Amphiphilic peptide D237
Genome Editor Cas Nuclease SpCas9
Guide BCM_ssAAV5-Sp_sgB This sgRNA targets the Ai9 and related transgenes
Guide SpyCas9 g-loxP2_C9 This sgRNA targets the mTmG transgene
Vector Viral Vector ssAAV5-sgA+sgB-U1A.GFP
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.
Model System Animal Ai9-SauSpyCas9 mouse Ai9-SauSpyCas9 is a version of Ai9 that has a single guide RNA sites on both sides of the STOP cassette. This is a Cre reporter allele that 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. Ai9 mice express robust tdTomato fluorescence following Cre-mediated recombination.
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
Vector Viral Vector ssAAV5-spCas9
Guide BCM_ssAAV5-Sp_sgA This sgRNA targets the Ai9 and related transgenes
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.
Experiment In Vivo Validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice 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
Delivery System Viral vector AAV See vector details
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.
Genome Editor Cas Nuclease SaCas9-Lagor
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_pTR_self comp 2xU6-Ai9 guides AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
Delivery System Nanoparticle 113-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.
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.
Genome Editor Cas Nuclease SauCas9
Guide Ai9 SaCas9 Guide B This gRNA targets the Ai9 and related transgenes
Guide STS159 (mTmG; Sp_t2:Sp_c20_mTmG) Targets mTmG and related reporter transgenes
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_141607  Invitrogen, Donkey anti-mouse alexafluor 488
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
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
Guide L2-modified This gRNA targets the Ai9 and related transgenes
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
Antibody AB_2286684  Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology
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.
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
Vector Viral Vector AAVcc47-Ai9-sgRNA1-CB-SaCas9 AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAV9-CMV-Cre AAV serotype 9 delivering CMV Cre Recombinase
Vector Viral Vector AAV9-Ai9-sgRNA2-CB-SaCas9 AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
Vector Viral Vector AAVcc47-CMV-SaCas9 AAVcc47 delivering CMV driven SaCas9
Guide sgAi9L This sgRNA targets the Ai9 and related transgenes
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
Model System Animal C57BL/6J mouse C57BL/6J WT mouse
Vector Viral Vector AAVcc47-CMV-Cre AAVcc47 delivering CMV Cre Recombinase
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.
Delivery System Nanoparticle 306-O12B Combinatorial library cationic lipid nanoparticles