利用TALEN技术高效制备TXNIP基因敲除小鼠模型  

Efficient preparation of a TXNIP knockout mouse model by transcription activator-like effector nucleases( TALEN)

作　　者：张欢欢[1] 刘楚新 马月[1] 肖丽萍 李飞达 应华忠[1] 刘欢 

机构地区：[1]浙江省医学科学院浙江省实验动物中心,杭州310013 [2]深圳华大基因研究院,深圳518083

出　　处：《中国比较医学杂志》2015年第6期9-13,共5页Chinese Journal of Comparative Medicine

基　　金：浙江省卫生高层次创新人才项目;浙江省科技厅院所专项(2014F10033)

摘　　要：目的利用显微注射技术注射敲除Txnip基因的TALEN mRNA获得TXNIP敲除小鼠。方法在线设计Txnip敲除位点,构建TALEN载体并在细胞水平验证剪切活性,体外转录TALENs成mRNA并通过显微注射技术注射到C57BL/6J小鼠受精卵,对F0代小鼠进行DNA水平鉴定获得TXNIP敲除小鼠。结果在Txnip第一外显子上设计了TALEN识别剪切位点,并在细胞水平验证具有剪切活性,注射mRNA到受精卵获得了4只敲除小鼠,其中2只Txnip发生移码突变,成功制备了TXNIP敲除小鼠。结论通过注射TALENs mRNA可以高效制备TXNIP敲除小鼠。Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease( TALEN) mRNAs. Methods TALEN knockout site recognizing Txnip was designed by tools on line,then constructed the vectors and assayed its cleavage activity at cellular level. TALEN mRNA was transcribed in vitro and microinjected into C57 BL /6J mouse zygotes. F0 mice were verified at DNA level with Bam HI and TXNIP-knockout mice were obtained. Results We designed and constructed TALENs which recognized and cut the first exon of Txnip,and got four TXNIP knockout mice,among which two were frameshift mutation,demonstrating that the TXNIP-knockout mice were generated by TALEN technique. Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

关 键 词：TALEN 显微注射 TXNIP 基因敲除 模型 小鼠 

分 类 号：R[医药卫生]