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CRISPR off-target analysis in genetically engineered rats and mice
- Anderson, Keith R;
- Haeussler, Maximilian;
- Watanabe, Colin;
- Janakiraman, Vasantharajan;
- Lund, Jessica;
- Modrusan, Zora;
- Stinson, Jeremy;
- Bei, Qixin;
- Buechler, Andrew;
- Yu, Charles;
- Thamminana, Sobha R;
- Tam, Lucinda;
- Sowick, Michael-Anne;
- Alcantar, Tuija;
- O’Neil, Natasha;
- Li, Jinjie;
- Ta, Linda;
- Lima, Lisa;
- Roose-Girma, Merone;
- Rairdan, Xin;
- Durinck, Steffen;
- Warming, Søren
Published Web Location
https://doi.org/10.1038/s41592-018-0011-5Abstract
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.
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