Abstract: Objective: A Golgi 73 (GP73) knockout mouse model was constructed and its phenotype was initially validated based on CRISPR/Cas9 technology.Methods: Non-homologous endjoining (NHEJ) was used to repair and introduce a mutation that caused a read frame shift in the GP73 gene, resulting in a loss of function.The sgRNA target sites were designed for exon 4 of the GP73 gene, and sgRNA was obtained by in vitro transcription.After mixing with mRNA encoding Cas9, F0 generation positive knockout mice were obtained by microinjection of fertilized eggs, and GP73 knockout homozygous mice (GP73^-/-mice) were obtained by breeding and genotype identification; real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect GP73 mRNA expression in liver, kidney, subcutaneous adipose tissue and brown adipose tissue, and serum GP73 protein level was detected by enzyme-linked immunosorbent assay (ELISA).Results: Compared with wildtype (WT) mice, GP73 mRNA and protein expressions in liver, kidney, subcutaneous adipose tissue as well as brown adipose tissue and serum GP73 protein levels of GP73^-/- mice decreased (all P＜ 0.05).Conclusion: GP73 knockout mouse models are successfully constructed based on CRISPR/Cas9 technology.