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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, 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  
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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Antibodies from Article