Frequency of off-targeting in genome edited pigs produced via direct injection of the CRISPR/Cas9 system into developing embryos

Published on Dec 1, 2019in BMC Biotechnology2.30
· DOI :10.1186/s12896-019-0517-7
Kayla Carey (VT: Virginia Tech), Junghyun Ryu5
Estimated H-index: 5
(VT: Virginia Tech)
+ 4 AuthorsKiho Lee6
Estimated H-index: 6
(VT: Virginia Tech)
Background The CRISPR/Cas9 system can effectively introduce site-specific modifications to the genome. The efficiency is high enough to induce targeted genome modifications during embryogenesis, thus increasing the efficiency of producing genetically modified animal models and having potential clinical applications as an assisted reproductive technology. Because most of the CRISPR/Cas9 systems introduce site-specific double-stranded breaks (DSBs) to induce site-specific modifications, a major concern is its potential off-targeting activity, which may hinder the application of the technology in clinics. In this study, we investigated off-targeting events in genome edited pigs/fetuses that were generated through direct injection of the CRISPR/Cas9 system into developing embryos; off-targeting activity of four different sgRNAs targeting RAG2, IL2RG, SCD5, and Ig Heavy chain were examined.
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