1. Efficient TALEN-mediated myostatin gene editing in goats
- Author
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Mengmin Zhu, Baoli Yu, Bin Shao, Yuguo Yuan, Shaozheng Song, Zhengqiang Qi, Rui Lu, Ting Zhang, Fei Mi, and Yong Cheng
- Subjects
0301 basic medicine ,Male ,Somatic cell ,Cell Culture Techniques ,Myostatin ,Genome ,03 medical and health sciences ,Genome editing ,Transcription (biology) ,Transcription Activator-Like Effector Nucleases ,Muscularity ,Animals ,Genetics ,Gene Editing ,Transcription activator-like effector nuclease ,biology ,Effector ,Goats ,0402 animal and dairy science ,04 agricultural and veterinary sciences ,Fibroblasts ,040201 dairy & animal science ,TALENs ,030104 developmental biology ,biology.protein ,Mutagenesis, Site-Directed ,Somatic cell nuclear transfer ,Female ,Mutational goat ,Nuclear transfer ,Developmental Biology ,Research Article - Abstract
Background Myostatin (MSTN) encodes a negative regulator of skeletal muscle mass that might have applications for promoting muscle growth in livestock. In this study, we aimed to test whether targeted MSTN editing, mediated by transcription activator-like effector nucleases (TALENs), is a viable approach to create myostatin-modified goats (Capra hircus). Results We obtained a pair of TALENs (MTAL-2) that could recognize and cut the targeted MSTN site in the goat genome. Fibroblasts from pedigreed goats were co-transfected with MTAL-2, and 272 monoclonal cell strains were confirmed to have mono- or bi-allelic mutations in MSTN. Ten cell strains with different genotypes were used as donor cells for somatic cell nuclear transfer, which produced three cloned kids (K179/MSTN−/−, K52-2/MSTN+/−, and K52-1/MSTN+/+). Conclusions The results suggested that the MTAL-2 could disrupt MSTN efficiently in the goat genome. The mutated somatic cells could be used to produce MSTN-site mutated goats without developmental disruption. Thus, TALENs is an effective method for accurate genome editing to produce site-modified goats.
- Published
- 2015