文献类型： 外文期刊
作者： Guo, Rihong 1 ; Wang, Huili 1 ; Meng, Chunhua 1 ; Gui, Hongbing 1 ; Li, Yinxia 1 ; Chen, Fang 1 ; Zhang, Chenjian 1 ; Zhang, Han 1 ; Ding, Qiang 1 ; Zhang, Jianli 1 ; Zhang, Jun 1 ; Qian, Yong 1 ; Zhong, Jifeng 1 ; Cao, Shaoxian 1 ;
作者机构： 1.Jiangsu Prov Engn Res Ctr Precis Anim Breeding, Nanjing 210014, Peoples R China
2.Jiangsu Acad Agr Sci, Inst Anim Sci, Nanjing 210014, Peoples R China
3.Minist Agr & Rural Affairs, Key Lab Crop & Anim Integrated Farming, Nanjing 210014, Peoples R China
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.
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