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Publication : CRISPR/Cas9-mediated knock-in of the murine Y chromosomal Sry gene.

First Author  Imaimatsu K Year  2018
Journal  J Reprod Dev Volume  64
Issue  3 Pages  283-287
PubMed ID  29657232 Mgi Jnum  J:308835
Mgi Id  MGI:6751786 Doi  10.1262/jrd.2017-161
Citation  Imaimatsu K, et al. (2018) CRISPR/Cas9-mediated knock-in of the murine Y chromosomal Sry gene. J Reprod Dev 64(3):283-287
abstractText  Mammalian zygote-mediated genome editing via the clustered regularly interspaced short palindromic repeats/CRISPR-associated endonuclease 9 (CRISPR/Cas9) system is widely used to generate genome-modified animals. This system allows for the production of loss-of-function mutations in various Y chromosome genes, including Sry, in mice. Here, we report the establishment of a CRISPR-Cas9-mediated knock-in line of Flag-tag sequences into the Sry locus at the C-terminal coding end of the Y chromosome (Y(Sry-flag)). In the F1 and successive generations, all male pups carrying the Y(Sry-flag) chromosome had normal testis differentiation and proper spermatogenesis at maturity, enabling complete fertility and the production of viable offspring. To our knowledge, this study is the first to produce a stable Sry knock-in line at the C-terminal region, highlighting a novel approach for examining the significance of amino acid changes at the naive Sry locus in mammals.
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