TALEN构建Fndc5基因敲除小鼠及初步分析

Constructtion of a Fndc5 knockout mouse model by TALEN-mediated DNA targeting

目的通过构建Fndc5基因敲除小鼠,为后续的研究提供动物模型。方法运用TALEN技术在Fndc5基因FNIII domain中造成缺失突变,并通过测序进行基因型鉴定。通过配对建立稳定遗传系并在mRNA和DNA水平鉴定出生小鼠基因型;对不同年龄段出生小鼠进行体重、血糖分析;通过q PCR确定Fndc5在肾脏、肝脏、大脑、肌肉、心脏等组织中的表达情况。结果成功构建并鉴定得到4种不同的Fndc5基因敲除小鼠品系;不同年龄段出生小鼠体重、血糖未见显著性差异;确定Fndc5在肌肉、心脏等组织中高表达。结论本实验在国际上成功构建了Fndc5基因敲除小鼠,并进行了初步分析,为深入研究Fndc5基因在体内中的功能提供了动物模型。

Objective To construct Fndc5 knockout mouse models and provide animal models for related studies in the future. Methods Indels were introduced into FNIII domain of Fndc5 gene by TALEN technology in mice,and genotypes were identified by sequencing. To set stable genetic system by pairing. Then at mRNA and DNA levels identified the genetype of born mice. At the same time the body weight and blood glucose of the mice at different ages were analyzed.Finally the Fndc5 expression in the kidney,liver,brain,muscles,heart and other organs was determined by q PCR.Results Four different Fndc5-KO lines were generated. The body weight and blood glucose of the mice at different agesshowed no significant differences. Finally high Fndc5 expressions in the muscles,heart and other organs were determined.Conclusions We Have for the first time successfully generated Fndc5 knockout（ KO） mouse model using TALEN mediated DNA targeting technique,and performed preliminary analysis. This Fndc5 knockout（ KO） mouse model provides a novel tool for further studies on the in vivo function of FNDC5.