%0 Journal Article
%T Genome editing using type I-E CRISPR-Cas3 in mice and rat zygotes.
%A Yoshimi K
%A Kuno A
%A Yamauchi Y
%A Hattori K
%A Taniguchi H
%A Mikamo K
%A Iida R
%A Ishida S
%A Goto M
%A Takeshita K
%A Ito R
%A Takahashi R
%A Takahashi S
%A Mashimo T
%J Cell Rep Methods
%V 4
%N 8
%D 2024 Aug 19
%M 39121862
暂无%R 10.1016/j.crmeth.2024.100833
%X The type I CRISPR system has recently emerged as a promising tool, especially for large-scale genomic modification, but its application to generate model animals by editing zygotes had not been established. In this study, we demonstrate genome editing in zygotes using the type I-E CRISPR-Cas3 system, which efficiently generates deletions of several thousand base pairs at targeted loci in mice with 40%-70% editing efficiency without off-target mutations. To overcome the difficulties associated with detecting the variable deletions, we used a newly long-read sequencing-based multiplex genotyping approach. Demonstrating remarkable versatility, our Cas3-based technique was successfully extended to rats as well as mice, even by zygote electroporation methods. Knockin for SNP exchange and genomic replacement with a donor plasmid were also achieved in mice. This pioneering work with the type I CRISPR zygote editing system offers increased flexibility and broader applications in genetic engineering across different species.