CRISPR off-target analysis in genetically engineered rats and mice

被引：176

作者：

Anderson, Keith R. ^{[1
]}

Haeussler, Maximilian ^{[2
]}

Watanabe, Colin ^{[3
]}

Janakiraman, Vasantharajan ^{[1
]}

Lund, Jessica ^{[1
]}

Modrusan, Zora ^{[1
]}

Stinson, Jeremy ^{[1
]}

Bei, Qixin ^{[1
]}

Buechler, Andrew ^{[1
]}

Yu, Charles ^{[1
]}

Thamminana, Sobha R. ^{[1
]}

Tam, Lucinda ^{[1
]}

Sowick, Michael-Anne ^{[4
]}

Alcantar, Tuija ^{[4
]}

O'Neil, Natasha ^{[4
]}

Li, Jinjie ^{[4
]}

Ta, Linda ^{[4
]}

Lima, Lisa ^{[4
]}

Roose-Girma, Merone ^{[1
]}

Rairdan, Xin ^{[4
]}

Durinck, Steffen ^{[1
]}

Warming, Soren ^{[1
]}

机构：

[1] Genentech Inc, Dept Mol Biol, San Francisco, CA 94080 USA

[2] Univ Calif Santa Cruz, Jack Baskin Sch Engn, Santa Cruz, CA 95064 USA

[3] Genentech Inc, Bioinformat & Computat Biol, San Francisco, CA 94080 USA

[4] Genentech Inc, Lab Anim Resources, San Francisco, CA 94080 USA

来源：

NATURE METHODS | 2018年 / 15卷 / 07期

关键词：

CAS9; NUCLEASES; DNA; SEQ; SPECIFICITY; CLEAVAGE; CELLS;

D O I：

10.1038/s41592-018-0011-5

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Despite widespread use of CRISPR, comprehensive data on the frequency and impact of Cas9-mediated off-targets in modified rodents are limited. Here we present deep-sequencing data from 81 genome-editing projects on mouse and rat genomes at 1,423 predicted off-target sites, 32 of which were confirmed, and show that high-fidelity Cas9 versions reduced off-target mutation rates in vivo. Using whole-genome sequencing data from ten mouse embryos, treated with a single guide RNA (sgRNA), and from their genetic parents, we found 43 off-targets, 30 of which were predicted by an adapted version of GUIDE-seq.

引用

页码：512 / +

页数：5

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