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CRISPR off-target analysis in genetically engineered rats and mice.

ABSTRACT: 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.

SUBMITTER: Anderson KR

PROVIDER: S-EPMC6558654 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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CRISPR off-target analysis in genetically engineered rats and mice.

Anderson Keith R KR Haeussler Maximilian M Watanabe Colin C Janakiraman Vasantharajan V Lund Jessica J Modrusan Zora Z Stinson Jeremy J Bei Qixin Q Buechler Andrew A Yu Charles C Thamminana Sobha R SR Tam Lucinda L Sowick Michael-Anne MA Alcantar Tuija T O'Neil Natasha N Li Jinjie J Ta Linda L Lima Lisa L Roose-Girma Merone M Rairdan Xin X Durinck Steffen S Warming Søren S

Nature methods 20180521 7

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 thei ...[more]

PMID: 29786090

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