Fundamental Toxicological Sciences

Paper Details

Fundamental Toxicological Sciences
Vol. 8 No. 5 October 28, 2021 p.161-167
Original Article
Diverse unintended on-target mutations induced by zygote genome-editing using CRISPR/Cas9 system
  • Toshime Igarashi (Division of Cellular & Molecular Toxicology, Center for Biological Safety & Research, National Institute of Health Sciences / toshime.igarashi@nihs.go.jp)
  • Satoshi Kitajima (Division of Cellular & Molecular Toxicology, Center for Biological Safety & Research, National Institute of Health Sciences / satoshi@nihs.go.jp)
Toshime Igarashi , Yukuto Yasuhiko , Ryuichi Ono , Erika Tachihara , Miki Uchiyama , Atsuya Takagi , Yu Takahashi , Makiko Kuwagata , Satoshi Kitajima
Division of Cellular & Molecular Toxicology, Center for Biological Safety & Research, National Institute of Health Sciences
Keywords: CRISPR/Cas9 system, Cas9 protein, Genome editing, Zygote electroporation, Knock-in, Mosaic
Abstracts

With the advent of the CRISPR/Cas9 system, genome editing in various fields is advancing. Unintended mutation in off-target regions is a major problem of genome editing using the CRISPR/Cas9 system, and it is being reviewed. However, we found a high frequency and various unintended mutations in the “on-target” region when we generated a “knock-in” mouse with point mutation using this technique to develop a supernumerary rib model. Additionally, an inserted sequence of unknown origin was observed. Furthermore, these mutations were transferred to the next generation, even if tandem knock-in or large deletions occurred. These strongly suggest that a proper selection that meets the purpose is essential when considering the safety of foods and medicines using the genome-editing technology.