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.
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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. |