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• W2788450735 abstract "Abstract Several groups recently reported coupling CRISPR/Cas9 perturbations and single cell RNA-seq as a potentially powerful approach for forward genetics. Here we demonstrate that vector designs for such screens that rely on cis linkage of guides and distally located barcodes suffer from swapping of intended guide-barcode associations at rates approaching 50% due to template switching during lentivirus production, greatly reducing sensitivity. We optimize a published strategy, CROP-seq, that instead uses a Pol II transcribed copy of the sgRNA sequence itself, doubling the rate at which guides are assigned to cells to 94%. We confirm this strategy performs robustly and further explore experimental best practices for CRISPR/Cas9-based single cell molecular screens." @default.
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• W2788450735 date "2018-01-29" @default.
• W2788450735 modified "2023-09-27" @default.
• W2788450735 title "On the design of CRISPR-based single cell molecular screens" @default.
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• W2788450735 doi "https://doi.org/10.1101/254334" @default.
• W2788450735 hasPublicationYear "2018" @default.
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