title: Silencing myostatin gene by RNAi in sheep embryos.
abstract: Myostatin (MSTN) gene is described as a negative regulator of the skeletal muscle growth. Controlling MSTN gene expression by genetic manipulation could accelerate the muscle growth and meat production of livestock animals. In the present study, several siRNAs targeting sheep MSTN gene were designed and their interfering efficiency was evaluated in vitro. The present study showed that one of the siRNAs, PSL1, could down-regulate the expression of MSTN significantly. PSL1 was ligated into lentivirus vector, GP-Supersilencing, to construct a siRNA expression lentivirus vector. Fibroblast cells were infected by lentivirus particles and positive cells were isolated by flow cytometry. Nucleus of the positive cell was transferred into enucleated oocytes of sheep. The present study showed that 99.4% of the sorted cells displayed green fluorescence. After enucleation of oocytes with microinjection, about 20% of reconstructed embryos can be developed into morulas, and strong green fluorescence could be observed using a fluorescence microscope. This method can be available to produce transgenic cell line for somatic cell nucleus transfer for transgenic animals.
authors.affiliation: Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Dayun
authors.initials: D
authors.lastname: Tang
authors.forename: Huabin
authors.initials: H
authors.lastname: Zhu
authors.forename: Jianmin
authors.initials: J
authors.lastname: Wu
authors.forename: Hanzhong
authors.initials: H
authors.lastname: Chen
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiaoliang
authors.initials: X
authors.lastname: Chen
authors.forename: Weihua
authors.initials: W
authors.lastname: Du
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
authors.forename: Xiukun
authors.initials: X
authors.lastname: Lin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D019976:Cloning, Organism; D004622:Embryo, Mammalian / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics; D020868:Gene Silencing; D005822:Genetic Vectors; D016086:Lentivirus; D055435:Myostatin / Q000096:biosynthesis* / Q000235:genetics; D053652:Nuclear Transfer Techniques; D034741:RNA, Small Interfering; D012756:Sheep
chemical_list: D055435:Myostatin; D034741:RNA, Small Interfering