134 results for AAV
134 results for AAV
AAV - Delivery System  - [In Vivo] [DCC, Delivery Systems Initiative] [Mouse] |
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing - Project  [Delivery Systems Initiative]![]()
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[Validation] Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing. - Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
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 - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVS1_site_01 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_02 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_05 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_08 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_10 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_13 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_14 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_04 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_09 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_11 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_06 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_03 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_12 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAVS1_site_07 - Guide  - [In Vitro] [Biological Systems] [Human] |
AAV9-GFP - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV9-mCherry - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV.pU1a-SpCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-Cre - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV9-Ai9-sgRNA1 + sgRNA2 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-CMV-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47_pTR_self comp 2xU6-Ai9 guides - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-SaCas9-Ai9 - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV9_pTR_self comp 2xU6-Ai9 guides - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-Ai9-sgRNA2-CB-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-Pcsk9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV9-Ai9-sgRNA2-CB-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV9-CMV-Cre - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-Ai9-sgRNA1 + sgRNA2 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc81-GFP - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAVcc47-CMV-Cre - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc47-mCherry - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAVcc84-GFP - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV9-CMV-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse]
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AAVcc47-Ai9-sgRNA1-CB-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV9-Ai9-sgRNA1-CB-SaCas9 - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1 - Vector  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
Ai9 SaCas9 Guide A - Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [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 Initiative] [Mouse] |
Cre Recombinase dose escalation study in Ai9 mice - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
*MOCK VIRUS TROPISM STUDY* For Toolkit demonstration purposes only - Experiment  - [In Vivo] [DCC] [Mouse] |
Ai9 SaCas9 Guide B - Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter) - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
Testing AAV5 for activation of tdTomato in HEK293T cells - Experiment  - [In Vitro] [Delivery Systems Initiative] [Human] |
Testing AAV5 for activation of tdTomato in mouse airway - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
Published procedures for AAV production, delivery, and tissue imaging.
Challis et al. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
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A novel human T cell platform to define biological adverse effects of genome editing - Experiment  - [In Vitro] [Biological Systems] [Human] |
SpCas9 - Genome Editor  - [In Vivo, In Vitro] [Delivery Systems Initiative, Biological Systems] [Human, Mouse] |
P3 Nucleofection Kit - Delivery System  - [In Vitro] [Biological Systems] [Human] |
CD4/CD8 Human Primary T cell - Model System  - [In Vitro] [Biological Systems] [Human] |
Cre recombinase - Genome Editor  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse]
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Procedure for Intratracheal (IT) delivery of AAV in mouse lung.
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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 Initiative] [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 Initiative] [Mouse] |
Testing AAV5 for activation of tdTomato in mouse airway club and ciliated cells - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
Testing virus region 4 (VR4) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
SaCas9-Lagor - Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse]
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Ai9 mouse - Model System  - [In Vivo] [Small Animal Testing Center (SATC), DCC, Delivery Systems Initiative] [Mouse]
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[Validation] Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain - Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
SauCas9 - Genome Editor  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
C57BL/6 mouse (Asokan study) - Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
Novel AAVs Engineered for Efficient and Noninvasive Cross-Species Gene Editing Throughout the Central Nervous System - Project  [Delivery Systems Initiative]![]() |
Deverman_Comprehensive_methods.pdf - Protocol  |
L1-modified - Guide  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse] |
SaCas9 - Genome Editor  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse] |
L3-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
BI28:AAV-GFAP-SaCas9-WPRE3-pA - Vector  - [In Vivo] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse] |
R1-modified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
R1-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
Testing gRNA sequence and gRNA scaffold modified in Ai9 mice. - Experiment  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
SaLoxP1-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
SaLoxP2-modified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
Ai9 mouse (BCM) - Model System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2 - Vector  - [In Vivo] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse] |
R2-modified - Guide  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Mouse] |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]Anti-RFP (RABBIT) Antibody |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000 |
TadA-8e V106W - Genome Editor  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
eVLP - Delivery System  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
L3-modifed - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
L1-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
L2-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]GFP (D5.1) XP Rabbit mAb antibody |
sgAi9R - Guide  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse] |
SaLoxP2-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
C57BL/6J mouse - Model System  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200 |
SaCas9 - Genome Editor  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
ssAAV5-sgB.saCas9 - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
ssAAV5-spCas9 - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
Selection of gRNA sequences and gRNA scaffold modification lead to improved editing of the Ai9 locus in vitro - Experiment  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
Ai9 mouse immortalized fibroblasts - Model System  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
R2-unmodified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
pH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1) - Vector  - [In Vitro] [Delivery Systems Initiative] [Human] |
HEK-293T with Ai9 transient reporter assay - Model System  - [In Vitro] [Delivery Systems Initiative] [Human] |
sgAi9L - Guide  - [In Vivo, In Vitro] [Delivery Systems Initiative] [Human, Mouse] |
pAAV.pU1a-SpCas9 - Vector  - [In Vitro] [Delivery Systems Initiative] [Human] |
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1) - Vector  - [In Vivo] [Delivery Systems Initiative] [Mouse] |
SaLoxP1-modified - Guide  - [In Vitro] [Delivery Systems Initiative] [Mouse] |
L2-modified - Guide  - [In Vivo, In Vitro] [Small Animal Testing Center (SATC), Delivery Systems Initiative] [Mouse] |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300 |
- Antibody  - [In Vivo] [Delivery Systems Initiative] [Mouse]Anti-RFP (Rabbit) Polyclonal Antibody |
Conklin_Fast-Seq_AAV.pdf - Protocol  |
BCM_ssAAV5-Sp_sgA - Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
Ai9-SauSpyCas9 mouse - Model System  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
Lipofectamine 3000 - Delivery System  - [In Vitro] [Genome Editors, Delivery Systems Initiative] [Human] |
[Validation] Validation for Gao Delivery Team: Testing ssAAV5 delivered intratracheally for editing activity in lung epithelia in Ai9 mice - Experiment  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
scAAV5-Cre-GFP - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
ssAAV5-sgA+sgB-U1A.GFP - Vector  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
BCM_ssAAV5-Sp_sgB - Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
SpCas9 - Genome Editor  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
BCM_ssAAV5-Sa_sgB - Guide  - [In Vivo] [Small Animal Testing Center (SATC)] [Mouse] |
134 results for AAV
Category | Type | Name | Description | Last Updated Date | View Associated.. |
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Delivery System | Viral vector | AAV | See vector details | ||
Project | Evolving High Potency AAV Vectors for Neuromuscular Genome Editing | Recombinant adeno-associated viruses (AAV) have emerged as safe and effective vectors for clinical gene therapy applications including systemic treatment of neuromuscular diseases such as Spinal Muscular Atrophy (SMA), Duchenne Muscular Dystrophy (DMD), and Giant Axonal Neuropathy (GAN) amongst others. However, genome editing in neuromuscular tissue, in particular, is challenging. The current proposal is on a comprehensive and innovative approach to evolve high potency AAV variants for systemic neuromuscular genome editing. | 2020-11-19 |
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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. | 2021-03-30 | |
Experiment | In Vivo | 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. | 2022-04-15 | |
Guide | AAVS1_site_01 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_02 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_05 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_08 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_10 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_13 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_14 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_04 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_09 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_11 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_06 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_03 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_12 | Targets AAVS1 safe harbor locus | |||
Guide | AAVS1_site_07 | Targets AAVS1 safe harbor locus | |||
Vector | Viral Vector | AAV9-GFP | AAV2/9 self complementary vector expressing GFP driven by CBh promoter | ||
Vector | Viral Vector | AAV9-mCherry | AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV.pU1a-SpCas9 | Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS | ||
Vector | Viral Vector | AAVcc47-Cre | AAV2/5 expressing Cre recombinase | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
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-CMV-SaCas9 | AAVcc47 delivering CMV driven SaCas9 | ||
Vector | Viral Vector | AAVcc47_pTR_self comp 2xU6-Ai9 guides | AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene | ||
Vector | Viral Vector | AAVcc47-SaCas9-Ai9 | AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
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-sgRNA2-CB-SaCas9 | AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Vector | Viral Vector | AAV-Pcsk9 | |||
Vector | Viral Vector | AAV9-Ai9-sgRNA2-CB-SaCas9 | AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus | ||
Vector | Viral Vector | AAV9-CMV-Cre | AAV serotype 9 delivering CMV Cre Recombinase | ||
Vector | Viral Vector | AAVcc47-Ai9-sgRNA1 + sgRNA2 | AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus | ||
Vector | Viral Vector | AAVcc81-GFP | AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Vector | Viral Vector | AAVcc47-CMV-Cre | AAVcc47 delivering CMV Cre Recombinase | ||
Vector | Viral Vector | AAVcc47-mCherry | AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter | ||
Vector | Viral Vector | AAVcc84-GFP | AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Vector | Viral Vector | AAV9-CMV-SaCas9 | AAV serotype 9 delivering CMV driven SaCas9 |
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Vector | Viral Vector | AAVcc47-Ai9-sgRNA1-CB-SaCas9 | AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus | ||
Vector | Viral Vector | AAV9-Ai9-sgRNA1-CB-SaCas9 | AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA | ||
Vector | Viral Vector | AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1 | AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence | ||
Guide | Ai9 SaCas9 Guide A | This gRNA targets the Ai9 and related transgenes | |||
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. | 2021-09-16 | |
Experiment | In Vivo | Cre Recombinase dose escalation study in Ai9 mice | A single stranded cmv cre cassette was packaged into AAV9 or AAVcc47 and injected intravenously in Ai9 mice. We injected n=3 at three different doses (1e10, 1e11, 1e12 vg) and harvested organs 4 weeks post injection. Fluorescence intensity in liver, heart, and skeletal muscle was quantified with tiff based images in Image J and neuronal transduction from each vector was quantified at the 1e12vg dose by counting the number of tdTomato+ neurons and number of NeuN+ cells from multiple sections and images. | 2021-09-16 | |
Experiment | In Vivo | *MOCK VIRUS TROPISM STUDY* For Toolkit demonstration purposes only | *MOCK STUDY* for demonstration purpose only. Data is fake. Five viral vector serotypes delivering Cre recombinase were tested by intravenous delivery into Ai9 mice and chacterized for biodistribution across 20 tissues by quantitative PCR and imaging. Cirulating IL-6 levels as well as cellular toxicity in the kidney were assessed 1-week post inoculation. | 2023-02-21 | |
Vector | Viral Vector | demo VV03-Cre | DEMO viral vector for demo purpose | ||
Vector | Viral Vector | demo VV05-Cre | DEMO viral vector for demo purpose | ||
Guide | Ai9 SaCas9 Guide B | This gRNA targets the Ai9 and related transgenes | |||
Experiment | In Vivo | Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to sgRNA ratio (CMV promoter) | A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=3) and AAVcc47 (n=3) by intravenous injection in Ai9 mice. A total dose of 3e12vg was injected into each mouse (1.5e12vg each vector mixed 1:1) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart. | 2021-09-16 | |
Experiment | In Vitro | Testing AAV5 for activation of tdTomato in HEK293T cells | AAV shuttle plasmids expressing SpCas9 and guide RNAs targeting the Ai9 transgene were tested in HEK293T cells by transient transfection. Both delivery and gene editing were detected by fluorescence. | 2020-10-20 | |
Experiment | In Vivo | Testing AAV5 for activation of tdTomato in mouse airway | AAV2/5 mediated gene editing in the mouse airway was tested by deliverying SpCas9 and guide RNAs targeting the Ai9 transgene in Ai9 transgenic mice. Viral delivery was detected by GFP expression and gene editing quantified by tdTomato activation | 2020-10-20 | |
Protocol | Deverman_AAV_production_and_administration_Nat Protocols-Challis_et_al_2019.pdf | Published procedures for AAV production, delivery, and tissue imaging. Challis et al. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3. | |||
Experiment | In Vitro | A novel human T cell platform to define biological adverse effects of genome editing | 110 guide RNAs and SpCas9 were transfected into human T-cells. Indel rates were measured by targeted amplicon deep sequencing. | 2020-12-09 | |
Genome Editor | Cas Nuclease | SpCas9 | HA-SV40NLS-SpCas9-SV40NLS | ||
Delivery System | Commercial Reagent | P3 Nucleofection Kit | Nuceleofection kit to be used with Lonza's nucleofection system. | ||
Model System | Cell | CD4/CD8 Human Primary T cell | CD4/CD8 Human Primary T cell | ||
Genome Editor | Recombinase | Cre recombinase | Cre recombinase delivered by AAV (see vector details) |
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Vector | Viral Vector | demo VV01-Cre | DEMO viral vector for demo purpose | ||
Vector | Viral Vector | demo VV02-Cre | DEMO viral vector for demo purpose | ||
Vector | Viral Vector | demo VV04-Cre | DEMO viral vector for demo purpose | ||
Protocol | Heaney-SATC_Gao-Validation_Intratracheal_Delivery_of_AAV_in_Mice.pdf | Procedure for Intratracheal (IT) delivery of AAV in mouse lung. | |||
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. | 2021-09-16 | |
Experiment | In Vivo | Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9 to sgRNA ratio (CMV promoter) | A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:4 ratio of cas9 to guide RNA (1e12vg of CMV Sacas9 vector and 3e12vg of the sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart. | 2021-09-16 | |
Experiment | In Vivo | 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. | 2021-09-21 | |
Experiment | In Vivo | Testing virus region 8 (VR8) mutant cross-species compatible Adeno Associated Viruses (ccAAVs) in mice. | C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding a GFP reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification. | 2020-11-19 | |
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. | 2020-11-19 | |
Genome Editor | Cas Nuclease | SaCas9-Lagor |
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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. |
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Experiment | In Vivo | Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to mouse brain | Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing. Editing detected and quantified in brain by generation of tdTomato fluorescent protein signal from Ai9 reporter mice | 2021-08-08 | |
Genome Editor | Cas Nuclease | SauCas9 | |||
Model System | Animal | C57BL/6 mouse (Asokan study) | |||
Project | Novel AAVs Engineered for Efficient and Noninvasive Cross-Species Gene Editing Throughout the Central Nervous System | This project aims to advance the NIH Somatic Cell Genome Editing Program’s objective to identify novel delivery technologies that enable genome editing in therapeutically relevant somatic cell populations. We will use proven virus engineering methods to develop new vehicles that can deliver genome editing machinery throughout the adult mammalian central nervous system. Accomplishing this objective would pave the road for applying gene editing, and gene therapy more broadly, to the study and treatment of neurological and psychiatric disorders. | 2021-04-17 | ||
Protocol | Deverman_Comprehensive_methods.pdf | Procedure for plasmid cloning, editing evaluation in fibroblast, AAV production and administration, tissue processing and IHC. | |||
Guide | L1-modified | This gRNA targets the Ai9 and related transgenes | |||
Genome Editor | Cas Nuclease | SaCas9 | |||
Guide | L3-unmodified | 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 | ||
Guide | R1-modified | This gRNA targets the Ai9 and related transgenes | |||
Guide | R1-unmodified | This gRNA targets the Ai9 and related transgenes | |||
Experiment | In Vivo | Testing gRNA sequence and gRNA scaffold modified in Ai9 mice. | 3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2 were codelivered intravenously to adult male and female Ai9 mice. Editing was assessed in brain sections 4 weeks later. | 2021-04-17 | |
Guide | SaLoxP1-unmodified | This gRNA targets the mTmG, Ai9 and related transgenes at two sites | |||
Guide | SaLoxP2-modified | This gRNA targets the mTmG, Ai9 and related transgenes at two sites | |||
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. | ||
Vector | Viral Vector | BI28:AAV-GFAP-NLS-GFP-WPRE-synpA-L1-R2 | Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds | ||
Guide | R2-modified | This gRNA targets the Ai9 and related transgenes |