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 qPCR. Results Four different Fndc5-KO lines were generated. The body weight and blood glucose of the mice at different ages showed 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.