Objective：To establish the S100A16 gene knockout mouse model，which can be used for the study on its biologic function. Methods：To establish the S100A16 gene knockout mouse model via Cre/loxP system. PCR was used to identify the genotype of the offspring，the expression level of S100A16 mRNA was detected by qRT-PCR，and expression of S100A16 protein was detected by Western blot. Results：S100A16 gene knockout mouse model has been successfully established. Heterozygous mice were successfully bred and reproduced. So far，gene knockout homozygous mice were not found. Conclusion：The S100A16 mouse could be a useful model for the researches on its function，especially in obesity and insulin resistance.