AAV
- Delivery System [Delivery Systems Initiative]
- [In Vivo]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
name -->
AAV
-->
AAV
-->
AAV
-->
AAV9-CMV-Cre
-->
AAV
-->
....AAV serotype 9 delivering CMV Cre Recombinase..See vector details..CAG-loxP-stop-loxP-tdtomato reporter
name -->
AAV
See vector details
|
AAVS1_site_05
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_05
-->
AAVS1_site_05
name -->
AAVS1_site_05
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_06
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_06
-->
AAVS1_site_06
name -->
AAVS1_site_06
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_14
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_14
-->
AAVS1_site_14
name -->
AAVS1_site_14
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_01
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_01
-->
AAVS1_site_01
name -->
AAVS1_site_01
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_03
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_03
-->
AAVS1_site_03
name -->
AAVS1_site_03
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_04
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_04
-->
AAVS1_site_04
name -->
AAVS1_site_04
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_11
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_11
-->
AAVS1_site_11
name -->
AAVS1_site_11
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_12
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_12
-->
AAVS1_site_12
name -->
AAVS1_site_12
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_08
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_08
-->
AAVS1_site_08
name -->
AAVS1_site_08
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_10
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_10
-->
AAVS1_site_10
name -->
AAVS1_site_10
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_09
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_09
-->
AAVS1_site_09
name -->
AAVS1_site_09
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_13
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_13
-->
AAVS1_site_13
name -->
AAVS1_site_13
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_02
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_02
-->
AAVS1_site_02
name -->
AAVS1_site_02
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAVS1_site_07
- Guide [Biological Systems]
- [In Vitro]
name -->
AAVS1_site_07
-->
AAVS1_site_07
name -->
AAVS1_site_07
-->
AAVS1
Targets AAVS1 safe harbor locus
|
AAV9-CMV-SaCas9
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-CMV-SaCas9
-->
AAV9_pTR_self comp 2xU6-Ai9 guides
AAV9-Ai9-sgRNA1 + sgRNA2
AAV9-CMV-SaCas9
AAVcc47_pTR_self comp 2xU6-Ai9 guides
-->
AAV serotype 9 delivering CMV driven SaCas9
-->
AAV
-->
..................................AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting
the Ai9 locus..AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector
) targeting Ai9 transgene..AAV serotype 9 delivering CMV driven SaCas9..AAVcc47 delivering u6 promoter
AAV serotype 9 delivering CMV driven SaCas9
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-Ai9-sgRNA1 + sgRNA2
-->
AAVcc47-Ai9-sgRNA1 + sgRNA2
AAVcc47-CMV-SaCas9
-->
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
-->
AAV
-->
..............AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus.
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-Ai9-sgRNA2-CB-SaCas9
-->
AAVcc47-Ai9-sgRNA2-CB-SaCas9
AAVcc47-Ai9-sgRNA1-CB-SaCas9
-->
AAV
AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
..This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
..This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
........This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
......This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
......This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
........This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
SaCas9 and U6 promoter driven gRNA..AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
AAV9-CMV-Cre
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV
symbol -->
AAV9-CMV-Cre
-->
AAV9-CMV-Cre
-->
AAV serotype 9 delivering CMV Cre Recombinase
-->
AAV
-->
........................................AAV serotype 9 delivering CMV Cre Recombinase..See vector details
AAV serotype 9 delivering CMV Cre Recombinase
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9_pTR_self comp 2xU6-Ai9 guides
-->
AAV9_pTR_self comp 2xU6-Ai9 guides
AAV9-CMV-SaCas9
-->
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting
-->
AAV
-->
..........AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector)
targeting Ai9 transgene..AAV serotype 9 delivering CMV driven SaCas9....CAG-loxP-stop-loxP-tdtomato reporter
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-Ai9-sgRNA1-CB-SaCas9
-->
AAVcc47-Ai9-sgRNA2-CB-SaCas9
AAVcc47-Ai9-sgRNA1-CB-SaCas9
-->
AAV
AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
........This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
..This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
symbol -->
AAV.pU1a-SpCas9
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-Ai9-sgRNA2-CB-SaCas9
-->
AAV9-Ai9-sgRNA2-CB-SaCas9
AAV9-Ai9-sgRNA1-CB-SaCas9
-->
AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
-->
AAV
-->
..........AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV serotype 9 delivering
AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus
|
AAVcc47-CMV-Cre
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-CMV-Cre
-->
AAVcc47-CMV-Cre
-->
AAV
AAVcc47 delivering CMV Cre Recombinase
|
AAVcc47-CMV-SaCas9
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-CMV-SaCas9
-->
AAVcc47-Ai9-sgRNA1 + sgRNA2
AAVcc47-CMV-SaCas9
-->
AAV
-->
..............AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus.
AAVcc47 delivering CMV driven SaCas9
|
AAVcc47-SaCas9-Ai9
- Vector
[Small Animal Testing Center (SATC)]
symbol -->
AAVcc47-SaCas9-Ai9
-->
AAVcc47-SaCas9-Ai9
-->
AAV
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
AAV2/9 expressing SaCas9 and single sgRNA under U6 promoter
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
......This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
........This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
SaCas9 and U6 promoter driven gRNA..AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-Ai9-sgRNA1 + sgRNA2
-->
AAV9-Ai9-sgRNA1 + sgRNA2
AAV9-CMV-SaCas9
-->
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
-->
AAV
-->
..................AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
..AAV serotype 9 delivering CMV driven SaCas9....CAG-loxP-stop-loxP-tdtomato reporter cassette which
AAV serotype 9 delivering u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47_pTR_self comp 2xU6-Ai9 guides
-->
AAV9-CMV-SaCas9
AAVcc47_pTR_self comp 2xU6-Ai9 guides
-->
AAV
-->
..........AAV serotype 9 delivering CMV driven SaCas9..AAVcc47 delivering u6 promoter driving sgRNA 1
AAVcc47 delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene
|
symbol -->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
-->
......This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
..This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
AAV9-mCherry
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-mCherry
-->
AAV9-mCherry
-->
AAV
AAV2/9 self complementary vector expressing Mcherry driven by CBh promoter
|
AAVcc47-Cre
- Vector
[Small Animal Testing Center (SATC)]
symbol -->
AAVcc47-Cre
-->
AAVcc47-Cre
-->
AAV
-->
Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
AAV2/5 expressing Cre recombinase
|
AAVcc47-mCherry
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc47-mCherry
-->
AAVcc47-mCherry
-->
AAV
AAV2/9 self complementary vector with capsid variant cc47 expressing Mcherry driven by CBh promoter
|
AAVcc84-GFP
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc84-GFP
-->
AAVcc84-GFP
-->
AAV
AAV2/9 self complementary vector with capsid variant cc84 expressing GFP driven by CBh promoter
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
........This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-Ai9-sgRNA1-CB-SaCas9
-->
AAV9-Ai9-sgRNA2-CB-SaCas9
AAV9-Ai9-sgRNA1-CB-SaCas9
-->
AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
-->
AAV
-->
..........AAV serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV serotype 9 delivering
AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus
|
AAVcc81-GFP
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAVcc81-GFP
-->
AAVcc81-GFP
-->
AAV
AAV2/9 self complementary vector with capsid variant cc81 expressing GFP driven by CBh promoter
|
symbol -->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3
-->
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold
-->
....This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3.gb
AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence
|
AAV9-GFP
- Vector
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
symbol -->
AAV9-GFP
-->
AAV9-GFP
-->
AAV
AAV2/9 self complementary vector expressing GFP driven by CBh promoter
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
Recombinant adeno-associated viruses (AAV) have emerged as safe and effective vectors for clinical gene
The current proposal is on a comprehensive and innovative approach to evolve high potency AAV variants
name -->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
Asokan Aravind
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.
|
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAVcc47-Cre
AAVcc47-SaCas9-Ai9
-->
Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery.
-->
AAV
-->
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see
AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
name -->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
Heaney Jason D
Quantification of CRISPR/Cas editing in liver and heart following custom AAV-mediated delivery. Detection of editing in non-target tissues.
|
Ai9 mouse
- Model System [Small Animal Testing Center (SATC), Delivery Systems Initiative]
- [In Vivo]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV
-->
AAV9-CMV-Cre
AAVcc47-CMV-SaCas9
AAV9-CMV-SaCas9
AAV.pU1a-SpCas9
AAVcc47-CMV-Cre
-->
AAV
-->
1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV
serotype 9 delivering CMV Cre Recombinase..Novel engineered AAV BI28 variant expressing S. aureus Cas9
HA-SV40NLS-SpCas9-SV40NLS..AAV serotype 9 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus..AAV
AAV2/5 expressing two sgRNAs under U6 promoter and eGFP..Novel engineered AAV BI28 variant expressing
delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene..AAV
-->
AAV BI28 (novel engineered variant)
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.
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV
-->
AAV9-CMV-Cre
AAVcc47-CMV-Cre
-->
AAV
-->
....................................................................................................AAV
....................................................................................................AAV
Asokan Aravind
A single stranded cmv cre cassette was packaged into AAV9 or AAVcc47 and injected intravenously in Ai9 mice. We injected n=3 at three different doses (1e10, 1e11, 1e12 vg) and harvested organs 4 weeks post injection. Fluorescence intensity in liver, heart, and skeletal muscle was quantified with tiff based images in Image J and neuronal transduction from each vector was quantified at the 1e12vg dose by counting the number of tdTomato+ neurons and number of NeuN+ cells from multiple sections and images.
|
-->
AAVS1_site_14
AAVS1_site_13
AAVS1_site_12
AAVS1_site_11
AAVS1_site_10
-->
AAVS1
Tsai Shengdar Q
110 guide RNAs and SpCas9 were transfected into human T-cells. Indel rates were measured by targeted amplicon deep sequencing.
|
P3 Nucleofection Kit
- Delivery System [Biological Systems]
- [In Vitro]
-->
AAVS1_site_14
AAVS1_site_13
AAVS1_site_12
AAVS1_site_11
AAVS1_site_10
-->
AAVS1
Nuceleofection kit to be used with Lonza's nucleofection system.
|
-->
AAVS1_site_14
AAVS1_site_13
AAVS1_site_12
AAVS1_site_11
AAVS1_site_10
-->
AAVS1
CD4/CD8 Human Primary T cell
|
SpCas9
- Genome Editor
[Biological Systems]
-->
AAVS1_site_14
AAVS1_site_13
AAVS1_site_12
AAVS1_site_11
AAVS1_site_10
-->
AAV.pU1a-SpCas9
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
-->
AAVS1
HA-SV40NLS-SpCas9-SV40NLS
|
-->
AAV
-->
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
Heaney Jason D
AAV5 encoding CRISPR/Cas editing machinery were delivered to the lungs of reporter mice by intratracheal instillation. After 4 weeks incubation, the mice were dissected and the lungs imaged for the presence of tdTomato fluorescence, indicating successful editing. Editing calculated by dividing the number of tdTomato+ red cells by the number of nuclei in each airway
|
scAAV5-Cre-GFP
- Vector
[Small Animal Testing Center (SATC)]
-->
AAV
-->
Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-Ai9-sgRNA1 + sgRNA2
AAVcc47-Ai9-sgRNA1 + sgRNA2
AAVcc47-CMV-SaCas9
AAV9-CMV-SaCas9
-->
single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV
-->
AAV
-->
single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV
calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart. ....................AAV
u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering CMV driven SaCas9..AAV
fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus.......................AAV
u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering CMV driven SaCas9..AAV
name -->
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 cas9 to
Asokan Aravind
A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=3) and AAVcc47 (n=3) by intravenous injection in Ai9 mice. A total dose of 3e12vg was injected into each mouse (1.5e12vg each vector mixed 1:1) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-Ai9-sgRNA1 + sgRNA2
AAVcc47-Ai9-sgRNA1 + sgRNA2
AAVcc47-CMV-SaCas9
AAV9-CMV-SaCas9
-->
single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV
-->
AAV
-->
single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV
calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart. ........................AAV
u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering CMV driven SaCas9..AAV
fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus...........................AAV
u6 promoter driving sgRNA 1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering CMV driven SaCas9..AAV
name -->
Comparing CRISPR/Cas9 gene editing efficiencies between AAV9 and AAVcc47 in Ai9 mice with a 1:3 Cas9
Asokan Aravind
A dual vector strategy was employed: one delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (both single stranded AAV cassettes). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:4 ratio of cas9 to guide RNA (1e12vg of CMV Sacas9 vector and 3e12vg of the sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.
|
SauCas9
- Genome Editor
[Small Animal Testing Center (SATC)]
-->
AAVcc47-SaCas9-Ai9
-->
AAV
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
|
Ai9 SaCas9 Guide B
- Guide [Small Animal Testing Center (SATC)]
- [In Vivo]
-->
AAVcc47-SaCas9-Ai9
-->
AAV
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
This gRNA targets the Ai9 and related transgenes
|
Ai9 SaCas9 Guide A
- Guide [Small Animal Testing Center (SATC)]
- [In Vivo]
-->
AAVcc47-SaCas9-Ai9
-->
AAV
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
This gRNA targets the Ai9 and related transgenes
|
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
Deverman Benjamin E
Reporter transgene activation by SaCas9 gRNA target and modified scaffold sequences by transient transfection in immortalized Ai9 mouse fibroblasts
|
Ai9 mouse (BCM)
- Model System [Small Animal Testing Center (SATC)]
- [In Vivo]
-->
AAVcc47-Cre
AAVcc47-SaCas9-Ai9
-->
AAV
-->
Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see
AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
Ai9 mouse has a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus.
|
Ai9-SauSpyCas9 mouse
- Model System [Small Animal Testing Center (SATC)]
- [In Vivo]
-->
AAV
-->
Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see
AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
Ai9-SauSpyCas9 is a version of Ai9 that has a single guide RNA sites on both sides of the STOP cassette. This is a Cre reporter allele that has a loxP-flanked STOP cassette preventing transcription of a CAG promoter-driven red fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus. Ai9 mice express robust tdTomato fluorescence following Cre-mediated recombination.
|
R2-modified
- Guide [Delivery Systems Initiative]
- [In Vivo, In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
and U6 promoter driven gRNA..Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein (GFAP) promoter ..AAV
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
-->
AAV BI28 (novel engineered variant)
This gRNA targets the Ai9 and related transgenes
|
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase
delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..This gRNA
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
Immortalized fibroblasts made from Ai9 (B6.Cg-Gt(ROSA)26Sor^tm9(CAG-tdTomato)Hze/J) mice
|
-->
Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing
-->
AAV
-->
Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing
................................This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
................................This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
name -->
Second site validation of Deverman delivery platform using engineered AAVs to deliver CRSIPR/Cas9 to
-->
AAV BI28 (novel engineered variant)
Heaney Jason D
Validation of delivery of AAV custom designed to cross the blood-brain barrier for CRISPR/Cas9 editing. Editing detected and quantified in brain by generation of tdTomato fluorescent protein signal from Ai9 reporter mice
|
SaCas9-Lagor
- Genome Editor
[Delivery Systems Initiative]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-Ai9-sgRNA2-CB-SaCas9
AAV9-Ai9-sgRNA1 + sgRNA2
AAVcc47-CMV-SaCas9
AAV9-Ai9-sgRNA1-CB-SaCas9
AAV9-CMV-SaCas9
-->
AAV
-->
..............................................................AAV serotype 9 delivering u6 promoter driving
1 + sgRNA2 targeting the Ai9 locus..AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV
serotype 9 delivering gRNA 2 + CB SaCas9 targeting the Ai9 locus..AAVcc47 delivering CMV driven SaCas9..AAV
delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene..AAV
serotype 9 delivering CMV driven SaCas9..AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus..AAV
|
Cre recombinase
- Genome Editor
[Small Animal Testing Center (SATC), Delivery Systems Initiative]
-->
AAVcc47-Cre
-->
Cre recombinase delivered by AAV (see vector details)
-->
AAV
-->
Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details
)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see vector
details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered by AAV (see
AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)..Cre recombinase delivered
by AAV (see vector details)..Cre recombinase delivered by AAV (see vector details)....AAV2/5 expressing
-->
Validation for Asokan Delivery Team: Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
Cre recombinase delivered by AAV (see vector details)
|
L2-modified
- Guide [Small Animal Testing Center (SATC), Delivery Systems Initiative]
- [In Vivo, In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
-->
AAV
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
and U6 promoter driven gRNA with modified scaffold sequence..Novel engineered AAV BI28 variant expressing
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
-->
AAV BI28 (novel engineered variant)
This gRNA targets the Ai9 and related transgenes
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9_pTR_self comp 2xU6-Ai9 guides
AAV9-CMV-SaCas9
AAVcc47_pTR_self comp 2xU6-Ai9 guides
-->
AAV
-->
calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart. ......................AAV
delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene..AAV
fluorescent protein variant (tdTomato) - all inserted into the Gt(ROSA)26Sor locus.........................AAV
delivering u6 promoter driving sgRNA 1 + sgRNA2 (self complementray vector) targeting Ai9 transgene..AAV
name -->
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
Asokan Aravind
A dual vector strategy was employed: one self complementary vector delivering two single guide RNAs targeting the Rosa26 locus and one delivering CMV driven SaCas9 (single stranded vector). This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=4) by intravenous injection in Ai9 mice. A total dose of 4e12vg was injected into each mouse and vectors mixed in a 1:1 ratio of cas9 to guide RNA (2e12vg of CMV Sacas9 vector and 2e12vg of the self complementary sgRNA vector) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.
|
SaCas9
- Genome Editor
[Small Animal Testing Center (SATC), Delivery Systems Initiative]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
AAV
-->
the mTmG, Ai9 and related transgenes at two sites..This gRNA targets the Ai9 and related transgenes ..AAV
shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven gRNA..AAV shuttle vector with
promoter driven SaCas9 and U6 promoter driven gRNA with modified scaffold sequence..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
-->
AAV BI28 (novel engineered variant)
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-Ai9-sgRNA2-CB-SaCas9
AAVcc47-Ai9-sgRNA2-CB-SaCas9
AAVcc47-Ai9-sgRNA1-CB-SaCas9
AAV9-Ai9-sgRNA1-CB-SaCas9
-->
AAV
-->
and heart. ........................AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV
CB SaCas9 targeting the Ai9 locus..AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus..AAV
26Sor locus...........................AAVcc47 delivering sgRNA 2 + CB SaCas9 targeting the Ai9 locus..AAV
CB SaCas9 targeting the Ai9 locus..AAVcc47 delivering sgRNA 1 + CB SaCas9 targeting the Ai9 locus..AAV
name -->
Comparing CRISPR/Cas9 gene editing efficencies between AAV9 and AAVcc47 in Ai9 mice with a 1:1 Cas9 to
Asokan Aravind
A dual vector strategy was employed: one delivering a single guide RNA and CB driven SaCas9, and another delivering the second guide RNA and CB driven SaCas9. This strategy was evaluted with both AAV9 (n=4) and AAVcc47 (n=5) by intravenous injection in Ai9 mice. A total dose of 2e12vg was injected into each mouse (1e12vg each vector mixed 1:1) and organs were harvested 4 weeks post injection. Editing efficency was determined by calculating percent TdTomato+ cells normalized to Dapi+ cells in liver and heart.
|
BI28:AAV-GFAP-SaCas9-WPRE3-pA
- Vector
[Small Animal Testing Center (SATC), Delivery Systems Initiative]
-->
Novel engineered AAV BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein
-->
AAV
-->
................................This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
-->
AAV BI28 (novel engineered variant)
Novel engineered AAV BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein (GFAP) promoter
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAVcc84-GFP
AAVcc81-GFP
AAV9-GFP
-->
AAV
Asokan Aravind
C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding a GFP reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
|
C57BL/6 mouse (Asokan study)
- Model System [Delivery Systems Initiative]
- [In Vivo]
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-mCherry
AAVcc47-mCherry
AAVcc84-GFP
AAVcc81-GFP
AAV9-GFP
-->
AAV
|
-->
Evolving High Potency AAV Vectors for Neuromuscular Genome Editing
-->
AAV9-mCherry
AAVcc47-mCherry
-->
AAV
Asokan Aravind
C57/BL6 mice (N=3) were injected intravenously at a dose of 5e13 vg/kg per mouse with a self-complementary AAV9 or ccAAV vector encoding an mCherry reporter. The biodistribution of of virus transduction was chacterized in various tissues and cell types by fluorescence imaging quantification.
|
-->
Procedure for Intratracheal (IT) delivery of AAV in mouse lung.
Procedure for Intratracheal (IT) delivery of AAV in mouse lung.
|
-->
3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2
-->
AAV
-->
3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2
sections 4 weeks later. ..............This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
ROSA)26Sor locus.................This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
-->
AAV BI28 (novel engineered variant)
Deverman Benjamin E
3e11 vg/mouse of AAV-BI28:GFAP-SaCas9-WPRE-pA and 3e11 vg/mouse of AAV-BI28:GFAP-NLS-GFP-U6-L1-U6-R2 were codelivered intravenously to adult male and female Ai9 mice. Editing was assessed in brain sections 4 weeks later.
|
L3-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
This gRNA targets the Ai9 and related transgenes
|
R1-modified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
This gRNA targets the Ai9 and related transgenes
|
SaLoxP1-modified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1
-->
This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP1.gb
This gRNA targets the mTmG, Ai9 and related transgenes at two sites
|
SaLoxP2-modified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2
-->
This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-LoxP2.gb
This gRNA targets the mTmG, Ai9 and related transgenes at two sites
|
L3-modifed
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
and U6 promoter driven gRNA..AAV shuttle vector with CMV promoter driven SaCas9 and U6 promoter driven
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L3.gb
This gRNA targets the Ai9 and related transgenes
|
L1-modified
- Guide [Small Animal Testing Center (SATC), Delivery Systems Initiative]
- [In Vivo, In Vitro]
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1
-->
AAV
-->
This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV BI28 variant expressing NLS-GFP
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
BI28 variant expressing S. aureus Cas9 driven by the glial fibrillary acidic protein (GFAP) promoter ..AAV
-->
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R1.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-R2.gb
AAV-CMV-SaCas9-U6-modified scaffold-Sa-L1.gb
-->
AAV BI28 (novel engineered variant)
This gRNA targets the Ai9 and related transgenes
|
name -->
Novel AAVs Engineered for Efficient and Noninvasive Cross-Species Gene Editing Throughout the Central
Deverman Benjamin E
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.
|
-->
Published procedures for AAV production, delivery, and tissue imaging.
Challis et al.
Systemic AAV vectors for widespread and targeted gene delivery in rodents.
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.
|
-->
Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP
-->
AAV
-->
................................This gRNA targets the Ai9 and related transgenes ..Novel engineered AAV
fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds..Novel engineered AAV
-->
AAV BI28 (novel engineered variant)
Novel engineered AAV BI28 variant expressing NLS-GFP driven by the glial fibrillary acidic protein (GFAP) promoter and dual sgRNAs with modified scaffolds
|
R2-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
This gRNA targets the Ai9 and related transgenes
|
SaLoxP1-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1
-->
This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP1.gb
This gRNA targets the mTmG, Ai9 and related transgenes at two sites
|
L1-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
This gRNA targets the Ai9 and related transgenes
|
L2-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
This gRNA targets the Ai9 and related transgenes
|
R1-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1
-->
This gRNA targets the Ai9 and related transgenes ..AAV shuttle vector with CMV promoter driven SaCas9
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-R1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L1.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L2.gb
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-L3.gb
This gRNA targets the Ai9 and related transgenes
|
SaLoxP2-unmodified
- Guide [Delivery Systems Initiative]
- [In Vitro]
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2
-->
This gRNA targets the mTmG, Ai9 and related transgenes at two sites..AAV shuttle vector with CMV promoter
-->
AAV-CMV-SaCas9-U6-gRNA-LoxP-Sa-LoxP2.gb
This gRNA targets the mTmG, Ai9 and related transgenes at two sites
|
-->
Procedure for plasmid cloning, editing evaluation in fibroblast, AAV production and administration, tissue
Procedure for plasmid cloning, editing evaluation in fibroblast, AAV production and administration, tissue processing and IHC.
|
-->
Published procedure for AAV composition measurement using fast-seq.
Maynard et al.
Published procedure for AAV composition measurement using fast-seq.
Maynard et al. Fast-Seq: A Simple Method for Rapid and Inexpensive Validation of Packaged Single-Stranded Adeno-Associated Viral Genomes in Academic Settings. Hum Gene Ther Methods
. 2019 Dec;30(6):195-205. doi: 10.1089/hgtb.2019.110.
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Cell Signaling Technology Cat# 2956
GFP (D5.1) XP Rabbit mAb antibody
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Rockland Cat# 600-401-379
Anti-RFP (RABBIT) Antibody
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Santa Cruz, SC-25555
Anti-CC10 (Rabbit) Polyclonal Antibody, dilution used 1:2,000
|
-->
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
Expresses codon-optimized SpCas9 in mammalian cells. HA-SV40NLS-SpCas9-SV40NLS
|
-->
AAV.pU1a-SpCas9
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
This sgRNA targets the Ai9 and related transgenes
|
-->
AAV.pU1a-SpCas9
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
This sgRNA targets the Ai9 and related transgenes
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Thermo Fisher Scientific, MA5-15257
Anti-RFP (Mouse) Monoclonal Antibody, dilution used 1:300
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Rockland, 600- 401-379
Anti-RFP (Rabbit) Polyclonal Antibody
|
-->
AAV.pU1a-SpCas9
H509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
-->
AAV
Other Id:
Santa Cruz, SC-23950
Anti-alpha Tubulin (Mouse) Monoclonal Antibody, dilution used 1:200
|
SpCas9
- Genome Editor
[Small Animal Testing Center (SATC)]
|
-->
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
HEK-293T cells transfected with an Ai9 inducible transgene reporter plasmid used to test gene editing activity by fluorescence
|
|
SaCas9
- Genome Editor
[Small Animal Testing Center (SATC)]
|
|
This sgRNA targets the Ai9 and related transgenes
|
Lipofectamine 3000
- Delivery System [Genome Editors]
- [In Vitro]
-->
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
Lipid nanoparticle
|
-->
PH509 AAVsc-u6-sgAI9L-U6-AI9R-U1A-EGFP (1)
AAV2/5 expressing SpyCas9. AAV2/5 expressing two sgRNAs under U6 promoter and eGFP
|
|
This gRNA targets the Ai9 and related transgenes
|
This sgRNA targets the Ai9 and related transgenes
|