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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, Aradhana CS, Abraham ND, Dao TJ, Thaseen S, Sairavi A, Gonzalez CU and Ploski JE.
Front. Mol. Neurosci. (2018) 13
Article

Plasmids from Article

ID Plasmid Purpose  
112138pJEP300-pAAV-CMV-MCS2-pA CMV driven Multi Cloning Site-2
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113676pJEP301-pAAV-CMV-MCS3-pACMV driven Multi Cloning Site-3
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113677pJEP302-pAAV-CMV-dSaCas9-KRAB-pAA CMV driven de-catalyzed SaCas9 fused to KRAB domain for inhibition of transcription in targeted region.
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113678pJEP303-pAAV-CMV-dSaCas9-VP64-pAA CMV driven de-catalyzed SaCas9 fused to VP64 domain for increased transcription in targeted region
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113679pJEP304-pAAV-EFS-dSaCas9-VP64-pAA EFS driven de-catalyzed SaCas9 fused to VP64 domain for increased transcription in targeted region
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113681pJEP305-pAAV-EFS-dSaCas9-KRAB-pAA EFS driven de-catalyzed SaCas9 fused to KRAB domain for inhibition of transcription in targeted region.
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113682pJEP306-pAAV-EFS-MCS3-pAEFS driven Multi Cloning Site-3.
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113684pJEP307-pAAV-EFS(No AgeI)-MCS3-pAEFS driven Multi Cloning Site-3 without the AgeI restriction enzyme site.
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113685pJEP308-pAAV-EFS-dSaCas9-VP64-Dio-pAAn EFS driven inverted de-catalyzed SaCas9 fused to the transcription activator VP64. dSaCas9-VP64 is floxed to render the system cre-dependent.
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113686pJEP309-pAAV-EFS-dSaCas9-KRAB-Dio-pAAn EFS driven inverted de-catalyzed SaCas9 fused to the transcription repressor KRAB. dSaCas9-KRAB is floxed to render the system cre-dependent.
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113687pJEP310-pAAV-Mecp2P-SaCas9-P2A-HAFLAGHA-KASH-pASaCas9 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.
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113688pJEP311-pAAV-EFS-SaCas9-P2A-HAFLAGHA-KASH-pASaCas9 driven by EFS. SaCas9 is followed by the self cleaving P2A sequence, several tags, and then the KASH transmembrane domain to enable FACS.
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113689pJEP312-pAAV-CMV-SaCas9-P2A-HAFLAGHA-KASH-pASaCas9 driven by CMV. SaCas9 is followed by the self cleaving P2A sequence, several tags, and then the KASH transmembrane domain to enable FACS
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113690pJEP313-pAAV-CMV-SaCas9-DIO-pAA CMV driven inverted SaCas9. SaCas9 is floxed to render the system cre-dependent.
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113691pJEP314-pAAV-U6SaCas9gRNA(SapI)-CMV-SaCas9-DIO-pAU6 driven SaCas9 gRNA expression cassette followed by a CMV driven inverted SaCas9. SaCas9 is floxed to render the system cre-dependent.
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113692pJEP315-pAAV-U6SaCas9gRNA(CREB)-CMV-SaCas9-DIO-pAU6 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.
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113693pJEP316-pAAV-U6SaCas9gRNA(SapI)-pA Empty CassetteU6 driven SaCas9 gRNA expression cassette without a gRNA. SapI can be used to clone in gRNAs.
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113694pJEP317-pAAV-U6SaCas9gRNA(SapI)-EFS-GFP- KASH-pAU6 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.
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113695pJEP318-pAAV-U6SaCas9gRNA(emx1sg1)-EFS-GFP-KASH-pAU6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1, followed by an EFS driven GFP-KASH in a separate reading frame.
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113696pJEP319-pAAV-U6SaCas9gRNA(emx1sg2)-EFS-GFP-KASH-pAU6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1, followed by an EFS driven GFP-KASH in a separate reading frame.
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113697pJEP320-pAAV-U6SaCas9gRNA(CREB3)_EFS-GFP-KASH-pAU6 driven SaCas9 gRNA expression cassette with a gRNA targeting CREB, followed by an EFS driven GFP-KASH in a separate reading frame.
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113698pJEP321-pAAV-FullU6TO-SaCas9gRNAi(SapI)-CMV-TetR-P2A-GFP-KASH-WPRE-shortPA Empty CassetteDox-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.
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113699pJEP322-pAAV-FullU6TO-SaCa9sgRNAi(emx1-sg1)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPADox-inducible U6 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1. Followed by an EFS driven GFP-KASH in a separate reading frame.
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113700pJEP323-pAAV-FullH1TO-SaCa9gRNAi(SapI)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPA Empty CassetteDox-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.
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113701pJEP324-pAAV-FullH1TO-SaCa9sgRNAi(emx1sg1)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPADox-inducible H1 driven SaCas9 gRNA expression cassette with a gRNA targeting EMX1. Followed by an EFS driven GFP-KASH in a separate reading frame.
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113702pJEP325-pAAV-FullH1TO-SaCa9sgRNAi(CREB)-CMV-TetR-2A-EGFP-KASH-WPRE-shortPADox-inducible H1 driven SaCas9 gRNA expression cassette with a gRNA targeting CREB. Followed by an EFS driven GFP-KASH in a separate reading frame.
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