High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening

被引：71

作者：

Gier, Rodrigo A. ^{[1
,2
,3
,6
]}

Budinich, Krista A. ^{[1
,2
,3
]}

Evitt, Niklaus H. ^{[1
,2
,3
,4
]}

Cao, Zhendong ^{[1
,2
,3
]}

Freilich, Elizabeth S. ^{[1
,2
,3
]}

Chen, Qingzhou ^{[1
,2
,3
]}

Qi, Jun ^{[5
]}

Lan, Yemin ^{[2
]}

Kohli, Rahul M. ^{[2
,4
]}

Shi, Junwei ^{[1
,2
,3
]}

机构：

[1] Univ Penn, Perelman Sch Med, Dept Canc Biol, Philadelphia, PA 19104 USA

[2] Univ Penn, Epigenet Inst, Perelman Sch Med, Dept Cell & Dev Biol, Philadelphia, PA 19104 USA

[3] Univ Penn, Abramson Family Canc Res Inst, Perelman Sch Med, Philadelphia, PA 19104 USA

[4] Univ Penn, Perelman Sch Med, Dept Med, Philadelphia, PA 19104 USA

[5] Harvard Med Sch, Dana Farber Canc Inst, Dept Med, Dept Canc Biol, Boston, MA 02215 USA

[6] Univ Penn, Dept Bioengn, Philadelphia, PA 19104 USA

来源：

NATURE COMMUNICATIONS | 2020年 / 11卷 / 01期

关键词：

LEUKEMIA; MAINTENANCE; CELLS; BRD9;

D O I：

10.1038/s41467-020-17209-1

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. Here, we present a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we can achieve high efficiency combinatorial genetic screening. We demonstrate the performance of our optimized AsCas12a (opAsCas12a) through double knockout screening against epigenetic regulators. This screen reveals synthetic sick interactions between Brd9&Jmjd6, Kat6a&Jmjd6, and Brpf1&Jmjd6 in leukemia cells.

引用

页数：9

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