Maximizing CRISPRi efficacy and accessibility with dual-sgRNA libraries and optimal effectors.
Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M
Elife. 2022 Dec 28;11:e81856. doi: 10.7554/eLife.81856. PubMed Article
Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M
Elife. 2022 Dec 28;11:e81856. doi: 10.7554/eLife.81856. PubMed Article
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 187239 | pJR98 | CR3 constant region - hU6 sgRNA promoter flanked by BsmBI sites |
| 187240 | pJR100 | Lentiviral sgRNA vector for Perturb-seq with mU6 sgRNA promoter, CR1 constant region with CS1 capture sequence in stem loop, and UCOE EF1alpha driving PURO-BFP marker expression |
| 187241 | pJR101 | Lentiviral sgRNA vector for Perturb-seq with mU6 sgRNA promoter, CR1 constant region with CS1 capture sequence in stem loop, and UCOE EF1alpha driving PURO-GFP marker expression |
| 187242 | pJR103 | Lentiviral sgRNA vector with mU6 sgRNA promoter, CR1 constant region, and UCOE EF1alpha driving PURO-BFP marker expression |
| 187243 | pJR104 | Lentiviral sgRNA vector with mU6 sgRNA promoter, CR1 constant region, and UCOE EF1alpha driving PURO-GFP marker expression |
| 187244 | pJR106 | Lentiviral sgRNA vector for CROP-seq with mU6 sgRNA promoter, CR1 constant region, and UCOE EF1alpha driving PURO-BFP marker expression |
| 187245 | pJR107 | Lentiviral sgRNA vector for CROP-seq with mU6 sgRNA promoter, CR1 constant region, and UCOE EF1alpha driving PURO-GFP marker expression |
| 187246 | hCRISPRi_dual_1_2 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 187247 | hCRISPRi_dual_3_4 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 187248 | hCRISPRi_dual_5_6 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 187249 | hCRISPRa_dual_1_2 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 187250 | hCRISPRa_dual_3_4 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 187251 | hCRISPRa_dual_5_6 | Ultra-compact, human genome-wide CRISPRi library targeting each gene with a single element encoding a dual sgRNA cassette. |
| 188765 | pHR-UCOE-SFFV-dCas9-XTEN80-KRAB(Kox1)-P2A-EGFP | dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) |
| 188766 | pHR-UCOE-EF1a-Zim3-dCas9-P2A-mCherry | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188767 | pHR-UCOE-SFFV-Zim3-dCas9-P2A-BFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188768 | pHR-UCOE-SFFV-Zim3-dCas9-P2A-Hygro | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188769 | pHR_UCOE-EF1a-dCas9-XTEN80-KRAB(Kox1)-IRES-mCherry | dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) |
| 188770 | pHR_UCOE-SFFV-dCas9-XTEN80-KRAB(Kox1)-IRES-mCherry | dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) |
| 188771 | pHR-UCOE-EFS-dCas9-XTEN80-KRAB(Kox1)-P2A-EGFP | dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) |
| 188772 | pHR-UCOE-CMV-dCas9-XTEN80-KRAB(Kox1)-P2A-EGFP | dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) |
| 188773 | pHR-UCOE-EF1a-Zim3-dCas9-loxP-P2A-EGFP-loxP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188774 | pHR-UCOE-SFFV-Zim3-dCas9-loxP-P2A-EGFP-loxP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188775 | pHR-UCOE-EF1a-Zim3-dCas9-BFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188776 | pHR-UCOE-SFFV-Zim3-dCas9-BFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188777 | pHR-UCOE-EF1a-Zim3-dCas9-P2A-BFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188778 | pHR-UCOE-EF1a-Zim3-dCas9-P2A-GFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188779 | pHR-UCOE-SFFV-Zim3-dCas9-P2A-mCherry | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS |
| 188898 | pHR-UCOE-SFFV-dCas9-P2A-EGFP | Human lentiviral vector for expression of dCas9 with a C-terminal HA-2xNLS and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element |
| 188899 | pHR-UCOE-SFFV-Zim3-dCas9-P2A-EGFP | dCas9 with an N-term Zim3 KRAB-NLS fusion, a C-term HA-2xNLS, and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element |
| 188900 | pHR-UCOE-SFFV-EGFP | Human lentiviral vector for expression of EGFP from a SFFV promoter with an upstream ubiquitous chromatin opening element |
| 188901 | pHR-UCOE-SFFV-SID4x-dCas9-XTEN80-KRAB(Kox1)-P2A-EGFP | dCas9 with an N-term Sid4x fusion, a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1) fusion, and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element |
| 188902 | pHR-UCOE-SFFV-dCas9-KRAB(Kox1)-MeCP2-P2A-EGFP | dCas9 with a C-term NLS-KRAB(Kox1)-MeCP2 fusion (no linker), and GFP linked by a P2A site from a SFFV promoter with an upstream ubiquitous chromatin opening element |