4.7 Article

Efficient TALEN-mediated gene knockin at the bovine Y chromosome and generation of a sex-reversal bovine

期刊

CELLULAR AND MOLECULAR LIFE SCIENCES
卷 78, 期 13, 页码 5415-5425

出版社

SPRINGER BASEL AG
DOI: 10.1007/s00018-021-03855-1

关键词

Bovine; Y chromosome; TALENs; Knockout; Knockin; Sry gene; Sex reversal

资金

  1. National Transgenic Breeding Project of China [2018ZX08007001]

向作者/读者索取更多资源

In this study, researchers efficiently knocked out four Y-linked genes and generated a sex-reversal bovine using gene editing and gene knockin technologies. The study demonstrated for the first time that the Sry gene is an important sex-determining gene in bovine, providing new insights into mammalian sex determination research and practical applications in agriculture.
Functional elucidation of bovine Y-chromosome genes requires available genome editing technologies. Meanwhile, it has yet to be proven whether the bovine Sry gene is the main or single factor involved in the development of the male phenotype in bovine. Here, we efficiently knocked out four Y-linked genes (Sry, ZFY, DDX3Y, and EIF2S3Y) in bovine fetal fibroblasts (BFFs) with transcription activator-like effector nucleases (TALENs) individually. Furthermore, we used TALEN-mediated gene knockin at the Sry gene and generated a sex-reversal bovine by somatic cell nuclear transfer (SCNT). The resulting bovine had only one ovary and was sterile. We demonstrate, for the first time, that the Sry gene is an important sex-determining gene in bovine. Our method lays a solid foundation for detecting the biology of the bovine Y chromosome, as it may provide an alternative biological model system for the study of mammalian sex determination, and new methods for the practical application in agricultural, especially for sex predetermination.

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