Efficient and Specific Generation of MSTN-Edited Hu Sheep Using C-CRISPR

文献类型： 外文期刊

第一作者： Guo, Rihong

作者： Guo, Rihong;Wang, Huili;Meng, Chunhua;Gui, Hongbing;Li, Yinxia;Chen, Fang;Zhang, Chenjian;Zhang, Han;Ding, Qiang;Zhang, Jianli;Zhang, Jun;Qian, Yong;Zhong, Jifeng;Cao, Shaoxian;Guo, Rihong;Wang, Huili;Meng, Chunhua;Li, Yinxia;Chen, Fang;Ding, Qiang;Zhang, Jianli;Zhang, Jun;Qian, Yong;Zhong, Jifeng;Cao, Shaoxian;Cao, Shaoxian

作者机构：

关键词： CRISPR; Cas9; Hu sheep; MSTN; double muscling phenotype; off-target effects

期刊名称：GENES （ 影响因子：3.5； 五年影响因子：3.9 ）

ISSN：

年卷期： 2023 年 14 卷 6 期

页码：

收录情况： SCI

摘要： Hu sheep, an indigenous breed in China known for its high fecundity, are being studied to improve their growth and carcass traits. MSTN is a negative regulator of muscle development, and its inactivation results in muscularity. The C-CRISPR system, utilizing multiple neighboring sgRNAs targeting a key exon, has been successfully used to generate genes for complete knockout (KO) monkeys and mice in one step. In this study, the C-CRISPR system was used to generate MSTN-edited Hu sheep; 70 embryos injected with Cas9 mRNA and four sgRNAs targeting exon 3 of sheep MSTN were transferred to 13 recipients. Out of 10 lambs born from five recipients after full-term pregnancies, nine had complete MSTN KO with various mutations. No off-target effects were found. These MSTN-KO Hu sheep showed a double-muscled (DM) phenotype, characterized by a higher body weight at 3 and 4 months old, prominent muscular protrusion, clearly visible intermuscular groves, and muscle hypertrophy. The molecular analysis indicated enhanced AKT and suppressed ERK1/2 signaling in the gluteus muscle of the edited Hu sheep. In conclusion, MSTN complete KO Hu sheep with a DM phenotype were efficiently and specifically generated using C-CRISPR, and the C-CRISPR method is a promising tool for farm animal breeding.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序