A Cas-embedding strategy for minimizing off-target effects of DNA base editors

被引：50

作者：

Liu, Yajing ^{[1
,2
]}

Zhou, Changyang ^{[2
]}

Huang, Shisheng ^{[1
,3
]}

Dang, Lu ^{[4
]}

Wei, Yu ^{[2
,3
]}

He, Jun ^{[1
,3
]}

Zhou, Yingsi ^{[2
]}

Mao, Shaoshuai ^{[2
]}

Tao, Wanyu ^{[1
,3
]}

Zhang, Yu ^{[1
]}

Yang, Hui ^{[2
]}

Huang, Xingxu ^{[1
]}

Chi, Tian ^{[1
]}

机构：

[1] ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China

[2] Chinese Acad Sci, Inst Neurosci,Shanghai Inst Biol Sci, State Key Lab Neurosci,Shanghai Res Ctr Brain Sci, Key Lab Primate Neurobiol,CAS Ctr Excellence Brai, Shanghai, Peoples R China

[3] Univ Chinese Acad Sci, Beijing, Peoples R China

[4] Guangzhou Med Univ, Canc Hosp & Inst, Guangzhou, Peoples R China

来源：

NATURE COMMUNICATIONS | 2020年 / 11卷 / 01期

基金：

美国国家科学基金会;

关键词：

RNA;

D O I：

10.1038/s41467-020-19690-0

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：

DNA base editors, typically comprising editing enzymes fused to the N-terminus of nCas9, display off-target effects on DNA and/or RNA, which have remained an obstacle to their clinical applications. Off-target edits are typically countered via rationally designed point mutations, but the approach is tedious and not always effective. Here, we report that the off-target effects of both A>G and C>T editors can be dramatically reduced without compromising the on-target editing simply by inserting the editing enzymes into the middle of nCas9 at tolerant sites identified using a transposon-based genetic screen. Furthermore, employing this Cas-embedding strategy, we have created a highly specific editor capable of efficient C>T editing at methylated and GC-rich sequences.

引用

页数：9

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