Department of Molecular Biology, Genentech, Inc., South San Francisco, CA, USA.
Jack Baskin School of Engineering, University of California, Santa Cruz, Santa Cruz, CA, USA.
Nat Methods. 2018 Jul;15(7):512-514. doi: 10.1038/s41592-018-0011-5. Epub 2018 May 21.
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.
尽管 CRISPR 得到了广泛应用，但关于 Cas9 介导的修饰啮齿动物中脱靶的频率和影响的综合数据仍然有限。在这里，我们展示了来自 81 个基因组编辑项目的深度测序数据，这些项目针对小鼠和大鼠基因组中的 1423 个预测脱靶位点，其中 32 个已经得到证实，并表明高保真 Cas9 版本降低了体内的脱靶突变率。使用来自十个用单向导 RNA（sgRNA）处理的小鼠胚胎及其遗传父母的全基因组测序数据，我们发现了 43 个脱靶位点，其中 30 个是通过 GUIDE-seq 的改编版本预测的。
