title: Targeted Editing of Myostatin Gene in Sheep by Transcription Activator-like Effector Nucleases.
abstract: Myostatin (MSTN) is a secreted growth factor expressed in skeletal muscle and adipose tissue that negatively regulates skeletal muscle mass. Gene knockout of MSTN can result in increasing muscle mass in sheep. The objectives were to investigate whether myostatin gene can be edited in sheep by transcription activator-like effector nucleases (TALENs) in tandem with single-stranded DNA oligonucleotides (ssODNs). We designed a pair of TALENs to target a highly conserved sequence in the coding region of the sheep MSTN gene. The activity of the TALENs was verified by using luciferase single-strand annealing reporter assay in HEK 293T cell line. Co-transfection of TALENs and ssODNs oligonucleotides induced precise gene editing of myostatin gene in sheep primary fibroblasts. MSTN gene-edited cells were successfully used as nuclear donors for generating cloned embryos. TALENs combined with ssDNA oligonucleotides provide a useful approach for precise gene modification in livestock animals.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China ; College of Life Sciences, Shihezi University, Shihezi, Xinjiang, 832003, China .
authors.forename: Xinxia
authors.initials: X
authors.lastname: Zhao
authors.affiliation: College of Life Sciences, Shihezi University, Shihezi, Xinjiang, 832003, China .
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Wujiafu
authors.initials: W
authors.lastname: Sai
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Qiao
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Jingliang
authors.initials: J
authors.lastname: Sheng
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, China.
authors.forename: Guozhong
authors.initials: G
authors.lastname: Li
authors.affiliation: College of Life Sciences, Shihezi University, Shihezi, Xinjiang, 832003, China .
authors.forename: Dawei
authors.initials: D
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Shihezi University, Shihezi, Xinjiang, 832003, China .
authors.forename: Shengwei
authors.initials: S
authors.lastname: Hu
keywords: Myostatin; Sheep Primary Fibroblasts; Targeted Gene Editing; Transcription Activator-like Effector Nucleases