Targeted Editing of Myostatin Gene in Sheep by Transcription Activator-like Effector Nucleases.
Targeted Editing of Myostatin Gene in Sheep by Transcription Activator-like Effector Nucleases.
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通过类转录激活因子效应核酸酶对绵羊肌肉生长抑制素基因进行靶向编辑
DOI：
10.5713/ajas.15.0041
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发表时间：
2016-03
期刊：
影响因子：
--
通讯作者：
Hu S
中科院分区：
文献类型：
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作者：
Zhao X;Ni W;Chen C;Sai W;Qiao J;Sheng J;Zhang H;Li G;Wang D;Hu S
肌生长抑制素（Myostatin，MSTs）是一种在骨骼肌和脂肪组织中表达的分泌性生长因子，其负调节骨骼肌质量。绵羊肌肉质量的增加可以通过敲除绵羊肌肉质量的基因来实现。目的研究转录激活因子样效应核酸酶（TALEN）与单链寡核苷酸（ssODNs）串联能否在绵羊体内编辑肌肉生长抑制素基因。我们设计了一对TALEN靶向绵羊MPEG4基因编码区中的高度保守序列。通过在HEK 293 T细胞系中使用荧光素酶单链退火报告基因测定来验证TALEN的活性。TALENs和ssODNs寡核苷酸共转染诱导绵羊原代成纤维细胞中肌肉生长抑制素基因的精确基因编辑。经基因编辑的细胞被成功用作产生克隆胚胎的核供体。TALEN与ssDNA寡核苷酸的组合为家畜动物中的精确基因修饰提供了有用的方法。
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.