Targeted Editing of Myostatin Gene in Sheep by Transcription Activator-like Effector Nucleases

• Main Authors: Xinxia Zhao, Wei Ni, Chuangfu Chen, Wujiafu Sai, Jun Qiao, Jingliang Sheng, Hui Zhang, Guozhong Li, Dawei Wang, Shengwei Hu
• Format: Article
• Language: English
• Published: Asian-Australasian Association of Animal Production Societies 2016-03-01
• Series: Asian-Australasian Journal of Animal Sciences
• Subjects: Transcription Activator-like Effector Nucleases; Myostatin; Targeted Gene Editing; Sheep Primary Fibroblasts
• Online Access: http://www.ajas.info/upload/pdf/ajas-29-3-413.pdf
• ISSN: 1011-2367; 1976-5517

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.