1. CRISPR off-target analysis in genetically engineered rats and mice
- Author
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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, and Warming, Søren
- Subjects
Biological Sciences ,Genetics ,Human Genome ,Biotechnology ,Good Health and Well Being ,Animals ,Cell Line ,Clustered Regularly Interspaced Short Palindromic Repeats ,Female ,Genetic Engineering ,Genomics ,Male ,Mice ,Multiplex Polymerase Chain Reaction ,RNA ,Rats ,Whole Genome Sequencing ,Technology ,Medical and Health Sciences ,Developmental Biology ,Biological sciences - 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.
- Published
- 2018