193 results for Epithelium
193 results for Epithelium
Cas9 ribonucleoprotein delivery targeted to kidney epitheliumProject [Delivery Systems, Collaborative Opportunity Fund, Biological Effects]Show Experiments (1) |
Podocyte-specific gene editing in human kidney organoidsExperiment - [In Vitro] [Delivery Systems, Collaborative Opportunity Fund, Biological Effects] |
Testing Shuttle Peptides ability to deliver GFP-NLS to airway epithelia.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
g-loxP2_C9Guide - [In Vivo] [Delivery Systems] [Mouse] |
sgAi9LGuide - [In Vivo, In Vitro] [Delivery Systems] [Human, Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]Anti-RFP (Rabbit) Polyclonal Antibody Show Experiments (1) |
BALB/c mouseModel System - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
SaCas9Genome Editor - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]GFP (D5.1) XP Rabbit mAb antibody Show Experiments (1) |
rabbit polyclonal antibody against club cell secretory protein, unconjugated, 07-623, EMD Show Experiments (1) |
Alt-R® S.p. Cas9 Nuclease V3Genome Editor - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (7)
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mTmG mouseModel System - [In Vivo] [Delivery Systems] [Mouse] |
ssAAV5-sgB.saCas9Vector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
BCM_ssAAV5-Sa_sgBGuide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cellsExperiment - [In Vivo] [Delivery Systems] [Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300 Show Experiments (1) |
S10Delivery System - [In Vivo, In Vitro] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Human, Mouse, Rhesus macaque]Show Experiments (12)
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ssAAV5-sgA+sgB-U1A.GFPVector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
BCM_ssAAV5-Sp_sgAGuide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Heaney-SATC_McCray-Validation_Intranasal Instillation in Mice ProtocolProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
rabbit polyclonal antibody to detect cytokeratin5+ basal cells, uncojugated, 905501, Biolegend Show Experiments (1) |
D237Delivery System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
ssAAV5-spCas9Vector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
SpyCas9 g-loxP2_C9Guide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai9 mouse (BCM)Model System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Heaney-SATC_McCray-Validation_Lung Inflation and Fixation ProtocolProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
SpCas9Genome Editor - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Heaney-SATC_Gao-Validation_Intratracheal Delivery of AAV in MiceProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai9-SauSpyCas9 mouseModel System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
anti-Wilms Tumor-1 (WT1) Show Experiments (1) |
Macaca mulatta (Rhesus Macaque)Model System - [In Vivo] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Rhesus macaque]Show Experiments (1) |
sg298Guide - [In Vivo, In Vitro] [Delivery Systems, Collaborative Opportunity Fund, Biological Effects] [Mouse]Show Experiments (3) |
Testing AAV5 for activation of tdTomato in mouse airwayExperiment - [In Vivo] [Delivery Systems] [Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]Anti-RFP (RABBIT) Antibody Show Experiments (1) |
Shuttle peptides enable in vivo gene editing with Cas9 and Cas12a RNP in mouse airway epitheliaExperiment - [In Vivo] [Delivery Systems] [Mouse] |
AsCas12a (IDT and Feldan Therapeutics)Genome Editor - [In Vivo] [Delivery Systems] [Mouse] |
g-loxPbot_C12aGuide - [In Vivo] [Delivery Systems] [Mouse] |
scAAV5-Cre-GFPVector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
SpCas9Genome Editor - [In Vivo, In Vitro] [Delivery Systems, Biological Effects] [Human, Mouse] |
AAV.pU1a-SpCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
sgAi9RGuide - [In Vivo, In Vitro] [Delivery Systems] [Human, Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000 Show Experiments (1) |
mouse monocolonal antibody against the acetylated α-tubulin, unconjugated, T6793, Sigma-Aldrich Show Experiments (1) |
[Validation] Independent validation for McCray Delivery Team: Delivery of CRISPR Ribonucleoproteins to Airway Epithelia Using Novel Amphiphilic PeptidesExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
D10Delivery System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Heaney_SATC Tissue Processing, Imaging and AnalysisProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CB promoter)Experiment - [In Vivo] [Delivery Systems] [Mouse] |
AAV9_pTR_self comp 2xU6-Ai9 guidesVector - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-Ai9-sgRNA2-CB-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
Chaikof-Associated Protocol 1_Retro-orbital administration and High throughput sequencingProtocol - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
[Validation] Independent validation of Chaikof delivery platform using virus-like particle (VLP) delivery to the mouse liverExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
ABE8eGenome Editor - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
eVLPDelivery System - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (3)
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AAV7-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
AAV8-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
sNLS-SpCas9-sNLSGenome Editor - [In Vivo, In Vitro] [Delivery Systems, Collaborative Opportunity Fund, Biological Effects, Animal Reporter and Testing Center] [Mouse]Show Experiments (6)
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sg298Guide - [In Vivo] [Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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113-O12BDelivery System - [In Vivo] [Delivery Systems] [Mouse] |
Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
AAV9-GFPVector - [In Vivo] [Delivery Systems] [Mouse] |
306-S10Delivery System - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (5)
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ABE8e-Cas9Genome Editor - [In Vivo, In Vitro] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Human, Rhesus macaque]Show Experiments (3)
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Testing new LNPs (lipid nanoparticles) for delivery of Fluc mRNA in adult miceExperiment - [In Vivo] [Delivery Systems] [Mouse] |
CleanCap® FlucGenome Editor - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
AAV9-Ai9-sgRNA1-CB-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
AAV9-CMV-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (3)
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ABE8eGenome Editor - [In Vitro] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Mouse, Rhesus macaque]Show Experiments (1) |
70. Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins.Publication - [In Vivo] [Delivery Systems] [Mouse]PII: S0092-8674(21)01484-7, PUBMED 35021064, PMC PMC8809250, DOI 10.1016/j.cell.2021.12.021 ABSTRACT: Methods to deliver gene editing agents in vivo as ribonucleoproteins could offer safety advantages over nucleic acid delivery approaches. We report the development and application of engineered DNA-free virus-like particles (eVLPs) that efficiently package and deliver base editor or Cas9 ribonucleoproteins. By engineering VLPs to overcome cargo packaging, release, and localization bottlenecks, we developed fourth-generation eVLPs that mediate efficient base editing in several primary mouse and h ... SCGE data tags...
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C57BL/6J mouseModel System - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (3) |
SpyCas9-3xNLSGenome Editor - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Human, Mouse]Show Experiments (7)
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AAV3b-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
AAVrh10-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
Cre Recombinase dose escalation study in Ai9 miceExperiment - [In Vivo] [Delivery Systems] [Mouse] |
[Validation] Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brainExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai14 gRNAGuide - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (3)
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Other Id:
RRID:AB_2209751
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Other Id:
RRID:AB_141637
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[Validation] Independent validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing.Experiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Enabling Nanoplatforms for Targeted in vivo Delivery of CRISPR/Cas9 Ribonucleoproteins in the Brain.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
Murray-SATC_Gong-Validaiton_Gong Study ProtocolProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Other Id:
Rockland Cat# 600-401-379
AB_2209751
Anti-RFP (RABBIT) Antibody |
[Validation] Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner earExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Gong_Intracranial Injection Procedure for MiceProtocol - [In Vivo] [Delivery Systems] [Mouse] |
Other Id:
ab150155
AB_2813835
Abcam, Donkey anti-rat alexa fluour 647 |
Other Id:
AB_141607
A21202
Invitrogen, Donkey anti-mouse alexafluor 488 |
R2-modifiedGuide - [In Vivo, In Vitro] [Delivery Systems] [Mouse] |
Deverman_Comprehensive MethodsProtocol - [In Vivo, In Vitro] [Delivery Systems] [Mouse] |
Deverman_Area Based Quantification of Editing Efficiency ProtocolProtocol - [In Vivo, In Vitro] [Delivery Systems] [Mouse] |
L1-modifiedGuide - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (3) |
Chaikof-Associated Protocol 2_Off-target editing AND Primers for sequencing analysisProtocol - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47_pTR_self comp 2xU6-Ai9 guidesVector - [In Vivo] [Delivery Systems] [Mouse] |
Testing new LNPs (lipid nanoparticles) for delivery of Cas9 mRNA/sgRNA in adult mouse cochleaExperiment - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-CreVector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
AAVrh8-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
Pcsk9 Exon 1 splice domainGuide - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse] |
AAV9-CMV-CreVector - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
[Validation] Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner earExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
CleanCap® Cas9 mRNA (5moU)Genome Editor - [In Vivo] [Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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Other Id:
RRID:AB_10711040
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AAVDelivery System - [In Vivo, In Vitro] [Delivery Systems, Biological Effects, AAV tropism] [Human, Mouse]Show Experiments (9)
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SaCas9Genome Editor - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
AAV+Focused UltrasoundDelivery System - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
Ai14 mouse (congenic)Model System - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (7)
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gRNA #8Guide - [In Vivo, In Vitro] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Rhesus macaque] |
Other Id:
AB_2532994
13-0300Â
Thermo-Fisher, Rat anti-GFAP |
306-O12B blankDelivery System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
L2-modifiedGuide - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse] |
AAV-Pcsk9Vector - [In Vivo] [Delivery Systems] [Mouse] |
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to sgRNA ratio (CMV promoter) and self complementary sgRNA vector.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
SaCas9-LagorGenome Editor - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (4)
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AAV9-Ai9-sgRNA1 + sgRNA2Vector - [In Vivo] [Delivery Systems] [Mouse] |
AAV9-Ai9-sgRNA2-CB-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-Ai9-sgRNA1-CB-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-CMV-SaCas9Vector - [In Vivo] [Delivery Systems] [Mouse] |
Chaikof-Associated Protocol 3_Serum ELISAsProtocol - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2Vector - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse] |
AAV Tropism projectExperiment - [In Vivo] [AAV tropism] [Mouse] |
AAV4-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
AAVrh74-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
STS204 (DNMT1; Sp_t2:Sp_c20_Dnmt1)Guide - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse] |
Other Id:
Vector Labs Cat# BA-1000
RRID:AB_2313606
Vector Labs Goat Anti-Rabbit IgG Biotinylated Cat. #BA-1000 |
RNP-NC-no ligandDelivery System - [In Vivo, In Vitro] [Delivery Systems, Collaborative Opportunity Fund, Biological Effects, Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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RNP-NC-RVGDelivery System - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Other Id:
RRID:AB_2532994
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Other Id:
RRID:AB_2813835
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306-O12BDelivery System - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (7)
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AAVcc84-GFPVector - [In Vivo] [Delivery Systems] [Mouse] |
SauCas9Genome Editor - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
FSD315Delivery System - [In Vivo, In Vitro] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Human, Rhesus macaque]Show Experiments (7)
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Testing preparation for independent validation at The Jackson Laboratory Small Animal Testing CenterExperiment - [In Vivo] [Delivery Systems] [Mouse] |
Other Id:
AB_1196615
RRID:AB_1196615
GFP (D5.1) XP Rabbit mAb antibody, Cell Signaling Technology Show Experiments (3)
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Murray-SATC_Gong-Validation Brain Injection in Mice ProtocolProtocol - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Other Id:
AB_141637
A21207
Invitrogen, Donkey anti-rabbit alexa fluor 594 |
Testing gRNA sequence and gRNA scaffold modified in Ai9 mice.Experiment - [In Vivo] [Delivery Systems] [Mouse] |
CleanCap® Cas9Genome Editor - [In Vivo, In Vitro] [Delivery Systems] [Mouse] |
Other Id:
AB_10015251
25-6790
Anti-Myosin VIIa antibody, Proteus Biosciences |
GFP-NLSGenome Editor - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
150. Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia.Publication - [In Vivo, In Vitro] [Delivery Systems] [Human, Mouse]PII: 10.1038/s41467-019-12922-y, PUBMED 31659165, PMC PMC6817825, DOI 10.1038/s41467-019-12922-y ABSTRACT: The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle ... SCGE data tags...
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H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)Vector - [In Vivo] [Delivery Systems] [Mouse] |
Antibody - [In Vivo] [Delivery Systems] [Mouse]Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200 Show Experiments (1) |
rabbit polyclonal antibody against the pulmonary-associated surfactant protein C (SPC), unconjugated, PA5-71680, Invitrogen Show Experiments (1) |
Cre recombinaseGenome Editor - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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Amphiphilic Peptides Deliver Base Editor RNPs to Rhesus Monkey AirwayExperiment - [In Vivo] [Delivery Systems, Genome Editors, Collaborative Opportunity Fund] [Rhesus macaque]Show Project |
mTmG mouse (congenic)Model System - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (7)
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[Validation] Independent validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 miceExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
BCM_ssAAV5-Sp_sgBGuide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai9 mouseModel System - [In Vivo] [Delivery Systems, AAV tropism, Animal Reporter and Testing Center] [Mouse]Show Experiments (11)
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On-target editing compared to 14 circle-seq nominated off-target sites of adenine base editor delivered by BE-eVLP vs AAV in the C57BL/6 liverExperiment - [In Vivo] [Delivery Systems] [Mouse]Show Project |
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter)Experiment - [In Vivo] [Delivery Systems] [Mouse] |
Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter)Experiment - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-Ai9-sgRNA1 + sgRNA2Vector - [In Vivo] [Delivery Systems] [Mouse] |
Pcsk9 adenine base editor efficiency in liver and nonliver tissueExperiment - [In Vivo] [Delivery Systems] [Mouse]Show Project |
TadA-8e V106WGenome Editor - [In Vivo] [Delivery Systems] [Mouse] |
AAV5-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
AAV6-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
AAV9-ZsGreen-CreVector - [In Vivo] [AAV tropism] [Mouse]Show Experiments (1) |
STS159 (mTmG; Sp_t2:Sp_c20_mTmG)Guide - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (4)
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Other Id:
Thermo Fisher Scientific Cat# G10362
RRID:AB_2536526
GFP Recombinant Rabbit Monoclonal Antibody, Thermo Fisher Scientific #G10362 Show Experiments (3)
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AAVcc47-CMV-CreVector - [In Vivo] [Delivery Systems] [Mouse]Show Experiments (1) |
RNP-NC-CPPDelivery System - [In Vivo, In Vitro] [Delivery Systems, Collaborative Opportunity Fund, Biological Effects, Animal Reporter and Testing Center] [Mouse]Show Experiments (6)
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Other Id:
RRID:AB_141607
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AAV9-mCherryVector - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc47-mCherryVector - [In Vivo] [Delivery Systems] [Mouse] |
AAVcc81-GFPVector - [In Vivo] [Delivery Systems] [Mouse] |
C57BL/6 mouse (Asokan study)Model System - [In Vivo] [Delivery Systems] [Mouse] |
179. Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing.Publication - [In Vivo] [Delivery Systems] [Mouse]PII: 10.1038/s41467-022-33745-4, PUBMED 36210364, PMC PMC9548504, DOI 10.1038/s41467-022-33745-4 ABSTRACT: Recombinant adeno-associated viral (AAV) vectors are a promising gene delivery platform, but ongoing clinical trials continue to highlight a relatively narrow therapeutic window. Effective clinical translation is confounded, at least in part, by differences in AAV biology across animal species. Here, we tackle this challenge by sequentially evolving AAV capsid libraries in mice, pigs and macaques. We discover a highly potent, cross-species compatible variant (AAV.cc47) that shows improved attrib ... SCGE data tags...
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AAVcc47-SaCas9-Ai9Vector - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai9 SaCas9 Guide AGuide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai9 SaCas9 Guide BGuide - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
[Validation] Independent validation of Sontheimer delivery platform using heavily modified guide RNAs complexed with Cas9 proteins to deliver CRISPR/Cas9 to mouse brainExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Ai14 mouseModel System - [In Vivo] [Delivery Systems] [Mouse] |
[Validation] Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brainExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
Other Id:
AB_10711040
ab104224Â
Abcam, mouse anti-NeuN |
[Validation] Independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner earExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
[Validation] Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner earExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
[Validation] Independent validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brainExperiment - [In Vivo] [Animal Reporter and Testing Center] [Mouse] |
SaCas9Genome Editor - [In Vivo, In Vitro] [Delivery Systems, Animal Reporter and Testing Center] [Mouse]Show Experiments (3) |
BI28:AAV-GFAP-SaCas9-WPRE3-pAVector - [In Vivo] [Delivery Systems, Animal Reporter and Testing Center] [Mouse] |
Other Id:
AB_2286684
sc-17320
Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology |
193 results for Epithelium
Category | Name | Description | Source | View Associated... |
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Project | Cas9 ribonucleoprotein delivery targeted to kidney epithelium |
Show Experiments (1) |
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Experiment | Podocyte-specific gene editing in human kidney organoids | Kidney organoids were derived from a human iPS cell line with Ai9 (tdTomato) fluorescence-on reporter knocked into the AAVS1 safe harbor locus. Intact kidney organoids were transfected with CRISPR ribonucleoprotein complexes with and without molecular targeting agent (MTA) specific for podocytes. Genome editing events were detected by induction of tdTomato from the Ai9 reporter. | ||
Experiment | 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. | ||
Guide | g-loxP2_C9 | This sgRNA targets the Ai9 and related transgenes | IDT | |
Guide | sgAi9L | This sgRNA targets the Ai9 and related transgenes | IDT | |
Antibody | Anti-RFP (Rabbit) Polyclonal Antibody |
Show Experiments (1) |
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Model System | 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). | The Jackson Laboratory |
Show Experiments (1) |
Genome Editor | SaCas9 | Vector Encoded (ssaav5-sga+sgb-u1a.gfp) | ||
Antibody | GFP (D5.1) XP Rabbit mAb antibody |
Show Experiments (1) |
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Antibody | RRID:AB_310759 | rabbit polyclonal antibody against club cell secretory protein, unconjugated, 07-623, EMD |
Show Experiments (1) |
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Genome Editor | Alt-R® S.p. Cas9 Nuclease V3 | Recombinant S. pyogenes Cas9 nuclease, purified from an E. coli strain expressing the nuclease. Contains nuclear localization sequence (NLS) and C-terminal 6-His tag. Provided in solution at 10 µg/µL. 100 µg of Cas9 nuclease = 610 pmol. | Integrated DNA Technologies |
Show Experiments (7)
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Model System | mTmG mouse | ROSAmT/mGÂ 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 | The Jackson Laboratory | |
Vector | ssAAV5-sgB.saCas9 | Gao Lab | ||
Guide | BCM_ssAAV5-Sa_sgB | This gRNA targets the Ai9 and related transgenes | Vector encoded | |
Experiment | Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cells | AAV2/5 mediated gene editing in the mouse airway was tested by deliverying SpCas9 and guide RNAs targeting the Ai9 transgene in Ai9 transgenic mice. Gene editing quantified by tdTomato activation and cell specific markers for club and ciliated cell types. | ||
Antibody | Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300 |
Show Experiments (1) |
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Delivery System | S10 | Shuttle peptide used to deliver reagents to airway epithelia | McCray Lab |
Show Experiments (12)
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Vector | ssAAV5-sgA+sgB-U1A.GFP | Gao Lab | ||
Guide | BCM_ssAAV5-Sp_sgA | This sgRNA targets the Ai9 and related transgenes | Vector encoded | |
Protocol | Heaney-SATC_McCray-Validation_Intranasal Instillation in Mice Protocol | Procedure for intranasal instillation of ribonucleoprotein/peptide complex in mice for delivery to the lung. | ||
Antibody | RRID:AB_2565050 | rabbit polyclonal antibody to detect cytokeratin5+ basal cells, uncojugated, 905501, Biolegend |
Show Experiments (1) |
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Delivery System | D237 | McCray Lab | ||
Vector | ssAAV5-spCas9 | Gao Lab | ||
Guide | SpyCas9 g-loxP2_C9 | This sgRNA targets the mTmG transgene | IDT | |
Model System | 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. | Baylor College of Medicine | |
Protocol | Heaney-SATC_McCray-Validation_Lung Inflation and Fixation Protocol | Procedure for mouse lung inflation and fixation. | ||
Genome Editor | SpCas9 | IDT | ||
Protocol | Heaney-SATC_Gao-Validation_Intratracheal Delivery of AAV in Mice | Procedure for Intratracheal (IT) delivery of AAV in mouse lung. | ||
Model System | Ai9-SauSpyCas9 mouse | Using CRISPR/Cas9 genome editing in mouse embryos, the existing Rosa-CAG-LSL-tdTomato-WPRE conditional allele Gt(ROSA)26Sortm9(CAG-tdTomato)Hze (commonly referred to as Ai9) was modified to duplicate the guide target sequences for S. pyogenes and S. aureus Cas9 found on the 3' end of the loxP-flanked stop cassette [SpyCas9 5'GTATGCTATACGAAGTTAT (PAM AGG); SauCas9 5'ACGAAGTTATATTAAGGGTT(PAM CCGGAT)] onto the 5' end of the stop cassette. With this modification, a single guide RNA for S. pyogenes or S. aureus Cas9 can be used to mediate deletion of the stop cassette by non-homologous end joining and activation of tdTomato expression. | Baylor College of Medicine | |
Antibody | RRID:AB_2043201 | anti-Wilms Tumor-1 (WT1) |
Show Experiments (1) |
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Model System | Macaca mulatta (Rhesus Macaque) | Tarantal Lab |
Show Experiments (1) |
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Guide | sg298 | This sgRNA targets the Ai9 and related transgenes at multiple sites | Synthego |
Show Experiments (3) |
Experiment | 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 | ||
Antibody | Anti-RFP (RABBIT) Antibody |
Show Experiments (1) |
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Experiment | 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 | AsCas12a (IDT and Feldan Therapeutics) | IDT and Feldan Therapeutics | ||
Guide | g-loxPbot_C12a | This sgRNA targets the Ai9 and related transgenes at two sites | IDT | |
Vector | scAAV5-Cre-GFP | Gao Lab | ||
Genome Editor | SpCas9 | HA-SV40NLS-SpCas9-SV40NLS | Vector Encoded | |
Vector | AAV.pU1a-SpCas9 | Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS | Addgene Gao Lab | |
Guide | sgAi9R | This sgRNA targets the Ai9 and related transgenes | IDT | |
Antibody | Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000 |
Show Experiments (1) |
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Antibody | RRID:AB_477585 | mouse monocolonal antibody against the acetylated α-tubulin, unconjugated, T6793, Sigma-Aldrich |
Show Experiments (1) |
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Experiment | [Validation] Independent 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 | D10 | McCray Lab | ||
Protocol | Heaney_SATC Tissue Processing, Imaging and Analysis | Procedure for tissue preparation, imaging and analysis. |
Show Experiments (4)
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Experiment | 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 | AAV9_pTR_self comp 2xU6-Ai9 guides | AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene | Asokan Lab | |
Vector | AAVcc47-Ai9-sgRNA2-CB-SaCas9 | AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus | Asokan Lab | |
Protocol | Chaikof-Associated Protocol 1_Retro-orbital administration and High throughput sequencing | This Protocol describes in vivo administration of BE-eVLP for Pcsk9 knockdown in liver, including sequencing endpoint. |
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Experiment | [Validation] Independent validation of Chaikof delivery platform using virus-like particle (VLP) delivery to the mouse liver | Virus-like particles carrying a CRISPR/Cas base editor and a guide RNA targeting the PSCK9 locus were injected i.v. into male and female mice. One week after injection, the mice were dissected, and genomic DNA isolated from a panel of organs. Targeted NGS was performed to evaluate the degree of editing at the PCSK9 locus in the liver (primary target), and non-target organs. Two potential off-target editing sites (OT6 and OT7) were also sequenced. | ||
Genome Editor | ABE8e | Catalytically impaired Cas fused to TadA deaminase | Liu Lab | |
Delivery System | eVLP | engineered virus like particles | Liu Lab |
Show Experiments (3)
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Vector | AAV7-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | AAV8-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Genome Editor | sNLS-SpCas9-sNLS | SpCas9 with N- and C-terminal SV40 NLS | Aldevron 9212-5MG |
Show Experiments (6)
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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 | Synthego |
Show Experiments (4)
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Antibody | RRID:AB_2762843 | Goat anti-chicken IgG-Alexa Fluor 488 | ||
Delivery System | 113-O12B | combinatorial library cationic lipid nanoparticles | Xu lab | |
Experiment | Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. | C57BL/6 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. | ||
Experiment | Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. | C57BL/6 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 | AAV9-GFP | AAV2/9 self complementary vector expressing GFP driven by CBh promoter | Asokan Lab | |
Delivery System | 306-S10 | combinatorial library cationic lipid nanoparticles | Xu lab |
Show Experiments (5)
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Genome Editor | ABE8e-Cas9 | A-to-G base editor | Feldan Therapeutics |
Show Experiments (3)
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Experiment | 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. | ||
Genome Editor | 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. | Trilink Biotechnologies |
Show Experiments (1) |
Vector | AAV9-Ai9-sgRNA1-CB-SaCas9 | AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus | Asokan Lab | |
Vector | AAV9-CMV-SaCas9 | AAV serotype 9 delivering CMV driven SaCas9 | Asokan Lab |
Show Experiments (3)
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Genome Editor | ABE8e | Tad8e deaminase fused to a nickase (D10A) spCas9 | David Liu Lab |
Show Experiments (1) |
Publication | Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins. | Methods to deliver gene editing agents in vivo as ribonucleoproteins could offer safety advantages over nucleic acid delivery approaches. We report the development and application of engineered DNA-free virus-like particles (eVLPs) that efficiently package and deliver base editor or Cas9 ribonucleoproteins. By engineering VLPs to overcome cargo packaging, release, and localization bottlenecks, we developed fourth-generation eVLPs that mediate efficient base editing in several primary mouse and human cell types. Using different glycoproteins in eVLPs alters their cellular tropism. Single injections of eVLPs into mice support therapeutic levels of base editing in multiple tissues, reducing serum Pcsk9 levels 78% following 63% liver editing, and partially restoring visual function in a mouse model of genetic blindness. In vitro and in vivo off-target editing from eVLPs was virtually undetected, an improvement over AAV or plasmid delivery. These results establish eVLPs as promising vehicles for therapeutic macromolecule delivery that combine key advantages of both viral and nonviral delivery. | ||
Model System | C57BL/6J mouse | C57BL/6J WT mouse | The Jackson Laboratory |
Show Experiments (3) |
Genome Editor | 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 | Sontheimer lab |
Show Experiments (7)
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Vector | AAV3b-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | AAVrh10-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | demo VV02-Cre | DEMO viral vector for demo purpose | Virus Company X | |
Experiment | 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. | ||
Experiment | [Validation] Independent validation of Gong delivery platform using RNP-loaded nanocages to deliver CRISPR/Cas9 to mouse brain | Delivery of CRISPR/Cas9 via ribonuclear protein (RNP) loaded nanocages (NC) to the brain in Ai14 mice by intracranial bilateral injection. Tissues were harvested 14 days after NC administeration. On-target and off-target editing was assessed. | ||
Guide | Ai14 gRNA | This sgRNA targets the Ai9 and related transgenes at multiple sites | IDT |
Show Experiments (3)
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Antibody | RRID:AB_2633281 | Goat anti-rabbit IgG-Alexa Fluor 555 | ||
Antibody | RRID:AB_2209751 | |||
Antibody | RRID:AB_141637 | |||
Experiment | [Validation] Independent 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 | 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. | ||
Protocol | Murray-SATC_Gong-Validaiton_Gong Study Protocol | Procedure for intracranial injection, immunofluorescence and imaging. | ||
Antibody | AB_2209751 | Anti-RFP (RABBIT) Antibody | ||
Experiment | [Validation] Repeat experiment of independent 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 | ||
Protocol | Gong_Intracranial Injection Procedure for Mice | Procedure for intracranial delivery to mouse brain. | ||
Antibody | AB_2813835 | Abcam, Donkey anti-rat alexa fluour 647 | ||
Antibody | AB_141607 | Invitrogen, Donkey anti-mouse alexafluor 488 | ||
Guide | R2-modified | This gRNA targets the Ai9 and related transgenes | Vector encoded | |
Protocol | Deverman_Comprehensive Methods | Procedure for plasmid cloning, editing evaluation in fibroblast, AAV production and administration, tissue processing and IHC. | ||
Protocol | Deverman_Area Based Quantification of Editing Efficiency Protocol | Procedure for non-IHC based image quantification of editing. | ||
Guide | L1-modified | This gRNA targets the Ai9 and related transgenes | Vector encoded |
Show Experiments (3) |
Protocol | Chaikof-Associated Protocol 2_Off-target editing AND Primers for sequencing analysis | This protocol describes in vivo adminstration and subsequent analysis of off-target editing in the liver. | ||
Vector | AAVcc47_pTR_self comp 2xU6-Ai9 guides | AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene | Asokan Lab | |
Experiment | 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 | AAVcc47-Cre | AAV2/5 expressing Cre recombinase | Asokan Lab | |
Vector | AAVrh8-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | demo VV03-Cre | DEMO viral vector for demo purpose | Virus Company X | |
Vector | demo VV04-Cre | DEMO viral vector for demo purpose | Virus Company X | |
Vector | demo VV05-Cre | DEMO viral vector for demo purpose | Virus Company X | |
Guide | Pcsk9 Exon 1 splice domain | Pcsk9 Exon 1 splice domain | Synthego | |
Vector | AAV9-CMV-Cre | AAV serotype 9 delivering CMV Cre Recombinase | Asokan Lab |
Show Experiments (1) |
Experiment | [Validation] Independent 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. Subset of mice were administered LNP via canalostomy injection compared to uninjected control mice. Tissues were harvested 6 days after LNP administeration. On-target and off-target editing was assessed. | ||
Genome Editor | 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. | Trilink Biotechnologies |
Show Experiments (4)
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Antibody | RRID:AB_10711040 | |||
Antibody | RRID:AB_2937041 | Chicken Polyclonal anti-NeuN antibody | ||
Delivery System | AAV | See vector details |
Show Experiments (9)
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Genome Editor | SaCas9 | Leong Lab |
Show Experiments (1) |
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Delivery System | AAV+Focused Ultrasound | See vector details | Leong Lab |
Show Experiments (1) |
Model System | 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. | The Jackson Laboratory |
Show Experiments (7)
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Antibody | RRID:AB_2762834 | Alexa fluor 555, Donkey anti-rabbit | ||
Antibody | RRID:AB_10563941 | Polyclonal rabbit anti-RFP antibody | ||
Guide | gRNA #8 | IDT | ||
Antibody | AB_2532994 | Thermo-Fisher, Rat anti-GFAP | ||
Delivery System | 306-O12B blank | Combinatorial library cationic lipid nanoparticles | Xu lab | |
Guide | L2-modified | This gRNA targets the Ai9 and related transgenes | Vector encoded | |
Vector | AAV-Pcsk9 | |||
Experiment | 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 | SaCas9-Lagor | William Lagor, Baylor College Of Medicine |
Show Experiments (4)
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Vector | AAV9-Ai9-sgRNA1 + sgRNA2 | AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus | Asokan Lab | |
Vector | AAV9-Ai9-sgRNA2-CB-SaCas9 | AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus | Asokan Lab | |
Vector | AAVcc47-Ai9-sgRNA1-CB-SaCas9 | AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus | Asokan Lab | |
Vector | AAVcc47-CMV-SaCas9 | AAVcc47 delivering CMV driven SaCas9 | Asokan Lab | |
Protocol | Chaikof-Associated Protocol 3_Serum ELISAs | This protocol describes in vivo adminstration and subsequent blood draws and ELISA on C57BL/6J mice. |
Show Experiments (1) |
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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 | Deverman Lab | |
Model System | B6 | C57BL/6J WT mouse | Baylor College of Medicine | |
Experiment | AAV Tropism project | Ten AAV serotypes delivering Cre recombinase were tested by intravenous delivery into Ai9 mice and chacterized for biodistribution across 20 tissues by quantitative PCR and imaging | ||
Vector | AAV4-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | AAVrh74-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Guide | STS204 (DNMT1; Sp_t2:Sp_c20_Dnmt1) | For endogenous locus | in house production | |
Antibody | RRID:AB_2313606 | Vector Labs Goat Anti-Rabbit IgG Biotinylated Cat. #BA-1000 | ||
Delivery System | 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. | Gong Lab |
Show Experiments (4)
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Delivery System | 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 | Gong Lab | |
Antibody | RRID:AB_2532994 | |||
Antibody | RRID:AB_2813835 | |||
Delivery System | 306-O12B | Combinatorial library cationic lipid nanoparticles | Xu lab |
Show Experiments (7)
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Vector | AAVcc84-GFP | AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter | Asokan Lab | |
Antibody | RRID:AB_10711040 | Mouse monoclonal [1B7] to NeuN - Neuronal Marker | ||
Genome Editor | SauCas9 | |||
Delivery System | FSD315 | Shuttle peptide used to deliver reagents to airway epithelia | McCray Lab |
Show Experiments (7)
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Experiment | Testing preparation for independent 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 | ||
Antibody | RRID:AB_1196615 | GFP (D5.1) XP Rabbit mAb antibody, Cell Signaling Technology |
Show Experiments (3)
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Protocol | Murray-SATC_Gong-Validation Brain Injection in Mice Protocol | Procedure for brain injection surgical procedure, pre- and post-operative care for mice. | ||
Antibody | AB_141637 | Invitrogen, Donkey anti-rabbit alexa fluor 594 | ||
Experiment | 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. | ||
Genome Editor | 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. | Trilink Biotechnologies | |
Antibody | AB_10015251 | Anti-Myosin VIIa antibody, Proteus Biosciences | ||
Genome Editor | GFP-NLS | Nuclear targeted GFP | Expressed From (Escherichia coli BL21DE3) |
Show Experiments (1) |
Publication | Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia. | The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSAmT/mG mice. We observe no evidence of short-term toxicity with a widespread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells. | ||
Vector | H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1) | AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP | Addgene Gao Lab | |
Antibody | Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200 |
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Antibody | RRID:AB_2717534 | rabbit polyclonal antibody against the pulmonary-associated surfactant protein C (SPC), unconjugated, PA5-71680, Invitrogen |
Show Experiments (1) |
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Genome Editor | Cre recombinase | Cre recombinase delivered by AAV (see vector details) |
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Experiment | Amphiphilic Peptides Deliver Base Editor RNPs to Rhesus Monkey Airway | We utilized novel amphiphilic shuttle peptides to deliver base editor ribonucleoprotein (RNP) into the airways to edit airway epithelial cells (CCR5 locus) of rhesus monkeys. The Cas9-ABE8e RNP and shuttle peptides S10 or FSD315 were aerosolized into the rhesus monkey trachea. Seven days later, tissues were obtained and dissected, and airway epithelia collected from the trachea, mainstem, and segmental bronchi using cytology brushes. DNA was extracted from epithelial cells and subjected to high-throughput sequencing. Using the FSD315 shuttle peptide and Cas9-ABE8e, we achieved a mean editing efficiency of 2.8% at the CCR5 locus in airway epithelial cells (range 0.02 – 5.3%) depending on the anatomic region sampled. To visualize the biodistribution of the RNPs within the respiratory tract and in specific cell types, we delivered a Cy5-fluorescent peptide fused to a nuclear localization signal (NLS-Cy5) using the S10 peptide. The lungs were obtained 1 and 2 hours post-delivery, fixed, and examined by microscopy. Epifluorescence and confocal microscopy documented an effective intra-nuclear delivery of NLS-Cy5 into epithelial cells throughout the respiratory tract, including large, medium, and small airways, and alveolar regions. Ongoing analyses will identify the NLS-Cy5-positive epithelial cell types using co-localization with fluorescently-labeled antibodies. In summary, using a rhesus monkey model, following a single delivery of adenine base editor RNPs to the airways in a clinically relevant manner we achieved up to 5.3% editing efficiency of the CCR5 locus in airway epithelia, a level considered therapeutically relevant in cystic fibrosis. |
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Model System | 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. | The Jackson Laboratory |
Show Experiments (7)
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Experiment | [Validation] Independent 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 | ||
Guide | BCM_ssAAV5-Sp_sgB | This sgRNA targets the Ai9 and related transgenes | Vector encoded | |
Model System | 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. | The Jackson Laboratory |
Show Experiments (11)
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Experiment | On-target editing compared to 14 circle-seq nominated off-target sites of adenine base editor delivered by BE-eVLP vs AAV in the C57BL/6 liver | On-target editing compared to off-target editing at 14 CIRCLE seq nominated sites in livers of an adenine base editor delivered by engineered virus-like particles (BE-eVLPs). Treated mice vs. untreated vs. AAV was assessed one week after systemic administration of BE-eVLPs or AAV-Pcsk9 to C57BL/6 mice. DNA sequencing reads containing A-T to G-C mutations within protospacer positions 4-10. |
Show Project |
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Experiment | 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. | ||
Experiment | 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 | AAVcc47-Ai9-sgRNA1 + sgRNA2 | AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus | Asokan Lab | |
Experiment | Pcsk9 adenine base editor efficiency in liver and nonliver tissue | Adenine base editing at the Pcsk9 exon 1 splice donor site in mouse heart, kidney, liver, lungs, muscle, and spleen was assessed one week after systemic administration of an adenine base editor delivered by engineered virus-like particles (BE-eVLPs) in C56BL/6 mice |
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Genome Editor | TadA-8e V106W | Catalytically impaired Cas fused to evolved TadA deaminase (TadA-8e V106W) | Chaikof Lab | |
Vector | AAV5-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | AAV6-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | AAV9-ZsGreen-Cre | AAV backbone with a bi-directional promoter driving zsGreen and Cre | Guangping Gao Lab |
Show Experiments (1) |
Vector | demo VV01-Cre | DEMO viral vector for demo purpose | Virus Company X | |
Guide | STS159 (mTmG; Sp_t2:Sp_c20_mTmG) | This sgRNA targets the mTmG transgene | in house production |
Show Experiments (4)
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Antibody | RRID:AB_2536526 | GFP Recombinant Rabbit Monoclonal Antibody, Thermo Fisher Scientific #G10362 |
Show Experiments (3)
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Vector | AAVcc47-CMV-Cre | AAVcc47 delivering CMV Cre Recombinase | Asokan Lab |
Show Experiments (1) |
Delivery System | 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 | Gong Lab |
Show Experiments (6)
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Antibody | RRID:AB_141607 | |||
Vector | AAV9-mCherry | AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter | Asokan Lab | |
Vector | AAVcc47-mCherry | AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter | Asokan Lab | |
Vector | AAVcc81-GFP | AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter | Asokan Lab | |
Model System | C57BL/6 mouse (Asokan study) | Unspecified | ||
Publication | Cross-species evolution of a highly potent AAV variant for therapeutic gene transfer and genome editing. | Recombinant adeno-associated viral (AAV) vectors are a promising gene delivery platform, but ongoing clinical trials continue to highlight a relatively narrow therapeutic window. Effective clinical translation is confounded, at least in part, by differences in AAV biology across animal species. Here, we tackle this challenge by sequentially evolving AAV capsid libraries in mice, pigs and macaques. We discover a highly potent, cross-species compatible variant (AAV.cc47) that shows improved attributes benchmarked against AAV serotype 9 as evidenced by robust reporter and therapeutic gene expression, Cre recombination and CRISPR genome editing in normal and diseased mouse models. Enhanced transduction efficiency of AAV.cc47 vectors is further corroborated in macaques and pigs, providing a strong rationale for potential clinical translation into human gene therapies. We envision that ccAAV vectors may not only improve predictive modeling in preclinical studies, but also clinical translatability by broadening the therapeutic window of AAV based gene therapies. | ||
Antibody | RRID:AB_141607 | Alexa fluor 488, Donkey anti-mouse | ||
Vector | AAVcc47-SaCas9-Ai9 | AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter | Asokan Lab | |
Guide | Ai9 SaCas9 Guide A | This gRNA targets the Ai9 and related transgenes | Vector encoded | |
Guide | Ai9 SaCas9 Guide B | This gRNA targets the Ai9 and related transgenes | Vector encoded | |
Experiment | [Validation] Independent validation of Sontheimer delivery platform using heavily modified guide RNAs complexed with Cas9 proteins to deliver CRISPR/Cas9 to mouse brain | Heavily modified guide RNAs complexed with Cas9 proteins are injected locally to mouse striatum to activate reporter gene (mGFP). Editing detected via DAB staining in coronal brain sections. | ||
Model System | 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. | The Jackson Laboratory | |
Experiment | [Validation] Independent 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_10711040 | Abcam, mouse anti-NeuN | ||
Experiment | [Validation] Independent 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 | [Validation] Repeat experiment of independent validation of Chen delivery platform using LNPs to deliver CRISPR/Cas9 to mouse inner ear | Repeat experiment of CRISPR/Cas9 delivery via bioreducible lipid nanoparticles (LNPs) to the inner ear in Ai14 mice. Subset of mice were administered LNP via canalostomy injection. Control mice were admininstered a blank LNP. Tissues were harvested 6 days after LNP administration. On-target editing was assessed by RFP (tdTomato) signal. | ||
Experiment | [Validation] Independent 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 | SaCas9 |
Show Experiments (3) |
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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 | Deverman Lab | |
Antibody | AB_2286684 | Sox-2 (Y-17) Antibody, Santa Cruz Biotechnology |