Insert title here
    登录 | 注册 | 期刊登录 | 收藏
 
 高级组合检索
遗传 2014, Vol. 36 (7) :685 -690 doi:10.3724/SP.J.1005.2014.0685 <<-上一篇 下一篇 ->>
引用: 刘丹, 杨宇, 严云勤,等.利用TALEN技术在牛胎儿成纤维细胞中敲除Myostatin基因[J].遗传,2014,36(7):685-690
利用TALEN技术在牛胎儿成纤维细胞中敲除Myostatin基因
杨翠翠, 佟慧丽, 马兴红, 杜巍, 刘丹, 杨宇, 严云勤
东北农业大学生命科学学院, 哈尔滨 150030
Myostatin knockout in bovine fetal fibroblasts by using TALEN
Cuicui Yang, Huili Tong, Xinghong Ma, Wei Du, Dan Liu, Yu Yang, Yunqin Yan
College of Life Science, Northeast Agricultural University, Harbin 150030, China
全文:     原文
摘要: 肌肉生长抑制素(Myostatin, MSTN)基因能够负向调节骨骼肌的生长和发育, 牛MSTN基因突变会出现“双肌”特征。文章利用转录激活因子样效应物核酸酶(TALENs)靶向敲除牛胎儿成纤维细胞的MSTN基因, 获得敲除MSTN基因的细胞系, 为制备MSTN基因敲除牛提供材料。构建一对MSTN基因的TALENs真核表达载体, 分别采用PEI转染试剂和电穿孔法进行牛胎儿成纤维细胞的转染, 测序结果表明TALEN技术可用于敲除牛MSTN基因, 利用T7核酸内切酶1(T7E1)检测其突变效率, 结果显示电穿孔转染的敲除效率为20.4%。通过有限稀释法, 共获得10个MSTN基因敲除的细胞克隆(包括MSTN-/-和MSTN+/-), 其靶位点敲除的碱基数分别是1~20不等, 部分会出现移码突变。出现移码突变的细胞系可用于MSTN基因敲除的转基因肉牛的制备。
关键词转染 敲除 TALENs
Abstract: Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and mutations of bovine MSTN gene can cause a “double-muscle” feature. To knock out MSTN gene in bovine fetal fibroblast by transcription activator-like effector nucleases (TALENs) and obtain MSTN knockout cell lines, we constructed one pair of MSTN-TALEN vector and transfected into bovine fetal fibroblast cells by PEI and electroporation. Sequencing results demonstrated that TALEN was available for MSTN knockout. T7 endonuclease 1 (T7E1) was used for the detection of mutation efficiency. The results indicated that knockout efficiency of electroporation transfection was 20.4%, and 10 MSTN+/- and MSTN-/- cell colonies were obtained via limiting dilution method. The deletion number of nucleotides ranged from 1 to 20, and some of them were frameshift mutation, which could provide the possibility in production of MSTN knockout cattle in the future.
Keywords: transfection knockout bovine TALENs
收稿日期:2013-12-16      发布日期:2014-06-23     
基金支持:国家转基因专项(编号:2011ZX08007-002)资助
Insert title here
更多服务: 作者自存档 | 投稿导航 | 全球期刊检索 | 查找审稿人
关于我们 | 隐私申明 | 免责声明 | 意见反馈 | 合作服务
Magsci ©  版权所有
Copyright © 2010 - 2013 Magtech. All Rights Reserved.