CRISPR/Cas9-mediated knock-in strategy at the Rosa26 locus in cattle fetal fibroblasts

ABSTRACT: The genetic modification of cattle has many agricultural and biomedical applications. However, random integration often leads to the unstable or differentially expression of the exogenous genes, which limit the application and development of transgenic technologies. Finding a safe locus suitable for site-specific insertion and efficient expression of exogenous genes is a good way to overcome these hurdles. In this study, we efficiently integrated three targeted vector into the cattle Rosa26 (cRosa26) by CRISPR/Cas9 technology in which EGFP was driven by CAG, EF1a, PGK and cRosa26 endogenous promoter respectively. The CRISPR/Cas9 knock-in system allows highly efficient gene insertion of different expression units at the cRosa26 locus. We also find that in the four cell lines, EGFP was stabl

SUBMITTER: Xie Y 

PROVIDER: S-EPMC9704577 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature