title: Production of myostatin-targeted goat by nuclear transfer from cultured adult somatic cells.
abstract: Myostatin, a member of the transforming growth factor-β family, acts as a negative regulator of skeletal muscle mass. In this study, myostatin-targeted caprine fibroblasts were obtained and subjected to SCNT to determine whether myostatin-knockout goats could be created. Fibroblasts from a 2-mo-old goat were transfected with a myostatin-targeted vector to prepare transgenic donor cells for nuclear transfer. After serum-starvation (for synchronization of the cell cycle), the percentage of transgenic fibroblasts in the G(0)/G(1) phase increased (66.2% vs. 82.9%; P < 0.05) compared with that in the control group, whereas the apoptosis rate and mitochondrial membrane potential were unaffected (P > 0.05). There were no significant differences between in vivo- and in vitro-matured oocytes as recipient cytoplasts for rates of fusion (86.5% vs. 78.4%), pregnancy (21.6% vs. 16.7%), or kidding (2.7% vs. 0%). One female kid from an in vivo-matured oocyte was born, but died a few hours later. Microsatellite analysis and polymerase chain reaction identification confirmed that this kid was genetically identical to the donor cells. Based on Western blot analysis, myostatin of the cloned kid was not expressed compared with that of nontransgenic kids. In conclusion, SCNT using myostatin-targeted 2-mo-old goat fibroblasts as donors has potential as a method for producing myostatin-targeted goats.
authors.affiliation: Center of Embryo Engineering and Technology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China.
authors.forename: Zheng-Rong
authors.initials: ZR
authors.lastname: Zhou
authors.forename: Bu-Shuai
authors.initials: BS
authors.lastname: Zhong
authors.forename: Ruo-Xin
authors.initials: RX
authors.lastname: Jia
authors.forename: Yong-Jie
authors.initials: YJ
authors.lastname: Wan
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Zhang
authors.forename: Yi-Xuan
authors.initials: YX
authors.lastname: Fan
authors.forename: Li-Zhong
authors.initials: LZ
authors.lastname: Wang
authors.forename: Ji-Hao
authors.initials: JH
authors.lastname: You
authors.forename: Zi-Yu
authors.initials: ZY
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D005347:Fibroblasts / Q000648:ultrastructure; D055786:Gene Knockout Techniques / Q000662:veterinary*; D005822:Genetic Vectors; D006041:Goats / Q000235:genetics*; D018482:Muscle, Skeletal / Q000254:growth & development; D055435:Myostatin / Q000235:genetics* / Q000502:physiology; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure; D011247:Pregnancy
chemical_list: D055435:Myostatin