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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. PubMed Article

Plasmids from Article

ID Plasmid Purpose
138143pLentiRNACRISPR_001 - hU6-DR_BsmBI-EFS-PguCas13b-NLS-2A-Puro-WPREExpresses 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
138144pLentiRNACRISPR_002 - hU6-DR_BsmBI-EFS-PguCas13b-NES-2A-Puro-WPREExpresses 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
138145pLentiRNACRISPR_003 - hU6-DR_BsmBI-EFS-PspCas13b-NLS-2A-Puro-WPREExpresses PspCas13b in mammalian cells, localized in the nucleus. For cloning of guide RNAs compatible with PspCas13b. Contains a 3' direct repeat. Clone using BsmBI.
138146pLentiRNACRISPR_004 - hU6-DR_BsmBI-EFS-PspCas13b-NES-2A-Puro-WPREExpresses 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
138147pLentiRNACRISPR_005 - hU6-DR_BsmBI-EFS-RfxCas13d-NLS-2A-Puro-WPREExpresses 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
138148pLentiRNACRISPR_006 - hU6-DR_BsmBI-EFS-RfxCas13d-NES-2A-Puro-WPREExpresses 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
138149pLentiRNACRISPR_007 - TetO-NLS-RfxCas13d-NLS-WPRE-EFS-rtTA3-2A-BlastExpresses RfxCas13d in mammalian cells for knockdown of target RNAs in combination with compatible crRNAs. Localized to the nucleus
138150pLentiRNAGuide_001 - hU6-RfxCas13d-DR1-BsmBI-EFS-Puro-WPREFor cloning of guide RNAs libraries compatible with RfxCas13d. Contains a 5' modified direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa
138151pLentiRNAGuide_002 - hU6-RfxCas13d-DR-BsmBI-EFS-Puro-WPREFor cloning of guide RNAs libraries compatible with RfxCas13d. Contains a 5' direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa
138152pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-HygroExpresses eGFP in mammalian cells, contains a 3' MCS. Localized to the nucleus

Antibodies from Article