On the design of CRISPR-based single-cell molecular screens.

Hill, Andrew J; McFaline-Figueroa, José L; Starita, Lea M; Gasperini, Molly J; Matreyek, Kenneth A; Packer, Jonathan; Jackson, Dana; Shendure, Jay et al. · Nat Methods · 2018

basic_science · Level V

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Abstract

Several groups recently coupled CRISPR perturbations and single-cell RNA-seq for pooled genetic screens. We demonstrate that vector designs of these studies are susceptible to ∼50% swapping of guide RNA-barcode associations because of lentiviral template switching. We optimized a published alternative, CROP-seq, in which the guide RNA also serves as the barcode, and here confirm that this strategy performs robustly and doubled the rate at which guides are assigned to cells to 94%.

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