Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modified Mice and Their Offspring. Issue 11 (1st November 2019)

• Record Type: Journal Article
• Authors: Dong, Yan; Li, Haimei; Zhao, Liang; Koopman, Peter; Zhang, Feng; Huang, Johnny X
• Is Part Of: G3. Volume 9:Issue 11(2019)
• Page Start: 3645
• Page End: 3651
• Publication Date: 2019-11-01
• Subjects: CRISPR-Cas9 -- Off-target -- Whole-genome sequencing -- Genome editing; Genetics -- Research -- Periodicals; Genomics -- Periodicals; Genetics; Genomics; Genes; Genetics -- Research; Genomics; Electronic journals; Periodical; Periodicals; Fulltext; Internet Resources; Periodicals; 572.8
• DOI: 10.1534/g3.119.400503
• Languages: English
• ISSNs: 2160-1836

Abstract

Abstract: The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. Here, we analyzed whole-genome sequencing (WGS) data from two knock-out (KO) mouse strains generated by using the CRISPR-Cas9 system targeting the Mmd and Paqr8 loci. A total of nine individuals were sequenced including two parents, four F1 offspring and three uninjected control mice. Using GATK and bcftools software, we identified two off-target events in the founder mice. The two CRISPR-Cas9-induced off-target events were predictable using Cas-OFFinder and were not passed on to the offspring that we investigated. In addition, our results indicated that the number of CRISPR-Cas9-induced mutations was not statistically distinguishable from the background de novo mutations (DNMs).