title: Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos.
abstract: Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and natural mutations can cause "double-muscling" trait in animals. In order to block the inhibiting effect of MSTN on muscle growth, we transferred zinc-finger nucleases (ZFN) which targeted sheep MSTN gene into cultured fibroblasts. Gene targeted colonies were isolated from transfected fibroblasts by serial dilution culture and screened by sequencing. Two colonies were identified with mono-allele mutation and one colony with bi-allelic deletion. Further, we introduced the MSTN-ZFN mRNA into sheep embryos by microinjection. Thirteen of thirty-seven parthenogenetic embryos were targeted by ZFN, with the efficiency of 35%. Our work established the technical foundation for generation of MSTN gene editing sheep by somatic cloning and microinjection ZFN into embryos.
authors.affiliation: Life Science and Technology, Xinjiang University, Urumqi, Xinjiang 830046, China.|Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Liqin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Yangsheng
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Life Science and Technology, Xinjiang University, Urumqi, Xinjiang 830046, China.|Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Wenrong
authors.initials: W
authors.lastname: Li
authors.affiliation: Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Jing
authors.initials: J
authors.lastname: An
authors.affiliation: Life Science and Technology, Xinjiang University, Urumqi, Xinjiang 830046, China.
authors.forename: Fuchun
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Life Science and Technology, Xinjiang University, Urumqi, Xinjiang 830046, China.|Key Laboratory of Genetics, Breeding and Reproduction of Grass-Feeding Animal, Ministry of Agriculture, Key Laboratory of Animal Biotechnology of Xinjiang, Urumqi, Xinjiang 830026, China.|Institute of animal biotechnology, Xinjiang Academy of Animal Science, Urumqi, Xinjiang 830026, China.
authors.forename: Mingjun
authors.initials: M
authors.lastname: Liu
keywords: Knockout; Myostatin; Sheep; Zinc-finger Nucleases