Massively parallel Cas13 screens reveal principles for guide RNA design.
Wessels HH, Mendez-Mancilla A, Guo X, Legut M, Daniloski Z, Sanjana NE
Nat Biotechnol. 2020 Jun;38(6):722-727. doi: 10.1038/s41587-020-0456-9. Epub 2020 Mar 16.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 138143 | pLentiRNACRISPR_001 - hU6-DR_BsmBI-EFS-PguCas13b-NLS-2A-Puro-WPRE | Expresses PguCas13b in mammalian cells, localized in the nucleus. For cloning of guide RNAs compatible with PguCas13b. Contains a 3' direct repeat. Clone using BsmBI. F overhang cacc. R overhang caac |
| 138144 | pLentiRNACRISPR_002 - hU6-DR_BsmBI-EFS-PguCas13b-NES-2A-Puro-WPRE | Expresses PguCas13b in mammalian cells, localized in the cytoplasm. For cloning of guide RNAs compatible with PguCas13b. Contains a 3' direct repeat. Clone using BsmBI. F overhang cacc. R overhang caac |
| 138145 | pLentiRNACRISPR_003 - hU6-DR_BsmBI-EFS-PspCas13b-NLS-2A-Puro-WPRE | Expresses PspCas13b in mammalian cells, localized in the nucleus. For cloning of guide RNAs compatible with PspCas13b. Contains a 3' direct repeat. Clone using BsmBI. |
| 138146 | pLentiRNACRISPR_004 - hU6-DR_BsmBI-EFS-PspCas13b-NES-2A-Puro-WPRE | Expresses PspCas13b in mammalian cells, localized in the cytoplasm. For cloning guide RNAs compatible with PspCas13b. Contains a 3' direct repeat. Clone using BsmBI. F overhang acac. R overhang caac |
| 138147 | pLentiRNACRISPR_005 - hU6-DR_BsmBI-EFS-RfxCas13d-NLS-2A-Puro-WPRE | Expresses RfxCas13d in mammalian cells, localized in the nucleus. For cloning of guide RNAs compatible with RfxCas13d. Contains a 5' direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa |
| 138148 | pLentiRNACRISPR_006 - hU6-DR_BsmBI-EFS-RfxCas13d-NES-2A-Puro-WPRE | Expresses RfxCas13d in mammalian cells, localized in the cytoplasm. For cloning of guide RNAs compatible with RfxCas13d. Contains a 5' direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa |
| 138149 | pLentiRNACRISPR_007 - TetO-NLS-RfxCas13d-NLS-WPRE-EFS-rtTA3-2A-Blast | Expresses RfxCas13d in mammalian cells for knockdown of target RNAs in combination with compatible crRNAs. Localized to the nucleus |
| 138150 | pLentiRNAGuide_001 - hU6-RfxCas13d-DR1-BsmBI-EFS-Puro-WPRE | For cloning of guide RNAs libraries compatible with RfxCas13d. Contains a 5' modified direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa |
| 138151 | pLentiRNAGuide_002 - hU6-RfxCas13d-DR-BsmBI-EFS-Puro-WPRE | For cloning of guide RNAs libraries compatible with RfxCas13d. Contains a 5' direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa |
| 138152 | pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro | Expresses eGFP in mammalian cells, contains a 3' MCS. Localized to the nucleus |