The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to Neurons in vivo and Regulated via Doxycycline and Cre-Recombinase.

Kumar N, Stanford W, de Solis C, Aradhana, Abraham ND, Dao TJ, Thaseen S, Sairavi A, Gonzalez CU, Ploski JE
Front Mol Neurosci. 2018 Nov 13;11:413. doi: 10.3389/fnmol.2018.00413. eCollection 2018. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
112138	pJEP300-pAAV-CMV-MCS2-pA	CMV driven Multi Cloning Site-2
113676	pJEP301-pAAV-CMV-MCS3-pA	CMV driven Multi Cloning Site-3
113677	pJEP302-pAAV-CMV-dSaCas9-KRAB-pA	A CMV driven de-catalyzed SaCas9 fused to KRAB domain for inhibition of transcription in targeted region.
113678	pJEP303-pAAV-CMV-dSaCas9-VP64-pA	A CMV driven de-catalyzed SaCas9 fused to VP64 domain for increased transcription in targeted region
113679	pJEP304-pAAV-EFS-dSaCas9-VP64-pA	A EFS driven de-catalyzed SaCas9 fused to VP64 domain for increased transcription in targeted region
113681	pJEP305-pAAV-EFS-dSaCas9-KRAB-pA	A EFS driven de-catalyzed SaCas9 fused to KRAB domain for inhibition of transcription in targeted region.
113682	pJEP306-pAAV-EFS-MCS3-pA	EFS driven Multi Cloning Site-3.
113684	pJEP307-pAAV-EFS(No AgeI)-MCS3-pA	EFS driven Multi Cloning Site-3 without the AgeI restriction enzyme site.
113685	pJEP308-pAAV-EFS-dSaCas9-VP64-Dio-pA	An EFS driven inverted de-catalyzed SaCas9 fused to the transcription activator VP64. dSaCas9-VP64 is floxed to render the system cre-dependent.
113686	pJEP309-pAAV-EFS-dSaCas9-KRAB-Dio-pA	An EFS driven inverted de-catalyzed SaCas9 fused to the transcription repressor KRAB. dSaCas9-KRAB is floxed to render the system cre-dependent.
113687	pJEP310-pAAV-Mecp2P-SaCas9-P2A-HAFLAGHA-KASH-pA	SaCas9 driven by the neuron specific promoter Mecp2P. SaCas9 is followed by the self cleaving P2A sequence, several tags, and then the KASH transmembrane domain to enable FACS.
113688	pJEP311-pAAV-EFS-SaCas9-P2A-HAFLAGHA-KASH-pA	SaCas9 driven by EFS. SaCas9 is followed by the self cleaving P2A sequence, several tags, and then the KASH transmembrane domain to enable FACS.
113689	pJEP312-pAAV-CMV-SaCas9-P2A-HAFLAGHA-KASH-pA	SaCas9 driven by CMV. SaCas9 is followed by the self cleaving P2A sequence, several tags, and then the KASH transmembrane domain to enable FACS
113690	pJEP313-pAAV-CMV-SaCas9-DIO-pA	A CMV driven inverted SaCas9. SaCas9 is floxed to render the system cre-dependent.
113691	pJEP314-pAAV-U6SaCas9gRNA(SapI)-CMV-SaCas9-DIO-pA	U6 driven SaCas9 gRNA expression cassette followed by a CMV driven inverted SaCas9. SaCas9 is floxed to render the system cre-dependent.
113692	pJEP315-pAAV-U6SaCas9gRNA(CREB)-CMV-SaCas9-DIO-pA	U6 SaCas9 gRNA expression cassette containing a gRNA targeting the CREB gene, followed by a CMV driven inverted SaCas9. SaCas9 is floxed to render the system cre-dependent.
113693	pJEP316-pAAV-U6SaCas9gRNA(SapI)-pA Empty Cassette	U6 driven SaCas9 gRNA expression cassette without a gRNA. SapI can be used to clone in gRNAs.
113694	pJEP317-pAAV-U6SaCas9gRNA(SapI)-EFS-GFP- KASH-pA	U6 driven SaCas9 gRNA expression cassette without a gRNA. Followed by an EFS driven GFP-KASH in a separate reading frame. SapI can be used to clone in gRNAs.
113695	pJEP318-pAAV-U6SaCas9gRNA(emx1sg1)-EFS-GFP-KASH-pA	U6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1, followed by an EFS driven GFP-KASH in a separate reading frame.
113696	pJEP319-pAAV-U6SaCas9gRNA(emx1sg2)-EFS-GFP-KASH-pA	U6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1, followed by an EFS driven GFP-KASH in a separate reading frame.
113697	pJEP320-pAAV-U6SaCas9gRNA(CREB3)_EFS-GFP-KASH-pA	U6 driven SaCas9 gRNA expression cassette with a gRNA targeting CREB, followed by an EFS driven GFP-KASH in a separate reading frame.
113698	pJEP321-pAAV-FullU6TO-SaCas9gRNAi(SapI)-CMV-TetR-P2A-GFP-KASH-WPRE-shortPA Empty Cassette	Dox-inducible U6 SaCas9 gRNA expression cassette without a gRNA. Followed by an EFS driven GFP-KASH in a separate reading frame. SapI can be used to clone in gRNAs.
113699	pJEP322-pAAV-FullU6TO-SaCa9sgRNAi(emx1-sg1)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPA	Dox-inducible U6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1. Followed by an EFS driven GFP-KASH in a separate reading frame.
113700	pJEP323-pAAV-FullH1TO-SaCa9gRNAi(SapI)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPA Empty Cassette	Dox-inducible H1 driven SaCas9 gRNA expression cassette without a gRNA. Followed by an EFS driven GFP-KASH in a separate reading frame. SapI can be used to clone in a gRNAs.
113701	pJEP324-pAAV-FullH1TO-SaCa9sgRNAi(emx1sg1)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPA	Dox-inducible H1 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1. Followed by an EFS driven GFP-KASH in a separate reading frame.
113702	pJEP325-pAAV-FullH1TO-SaCa9sgRNAi(CREB)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPA	Dox-inducible H1 driven SaCas9 gRNA expression cassette with a gRNA targeting CREB. Followed by an EFS driven GFP-KASH in a separate reading frame.