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

被引：0

作者：

Hill, Andrew J. ^{[1
]}

McFaline-Figueroa, Jose L. ^{[1
]}

Starita, Lea M. ^{[1
]}

Gasperini, Molly J. ^{[1
]}

Matreyek, Kenneth A. ^{[1
]}

Packer, Jonathan ^{[1
]}

Jackson, Dana ^{[1
]}

Shendure, Jay ^{[1
,2
]}

Trapnell, Cole ^{[1
]}

机构：

[1] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA

[2] Howard Hughes Med Inst, Seattle, WA 98195 USA

来源：

NATURE METHODS | 2018年 / 15卷 / 04期

关键词：

GENETIC SCREENS; EXPRESSION; CIRCUITS; GENOME; SEQ; P53;

D O I：

10.1038/NMETH.4604

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 similar 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%.

引用

页码：271 / +

页数：10

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