Objective：The aim of this study was to construct zinc transporter 8（ZnT8）chimeric antigen receptor（CAR）expression vector，so as to provide experimental basis for studying the role of antigen-specific regulatory T cells（Tregs）in the autoimmunity of type 1 diabetes. Methods：PLVX-EGFP-ZnT8 scfv lentivector was constructed by molecular cloning technique to produce ZnT8 scfv-CAR lentivirus；the expression of green fluorescent protein（GFP）in Tregs was detected by flow cytometry to determine the efficiency of lentivirus；the proliferation of Tregs was evaluated by cell counting；the expression of CD4，CD25，Foxp3 and Helios in proliferated cells were detected by flow cytometry. Results：The PLVX-EGFP-ZnT8 scfv lentivector was successfully constructed. The titer of concentrated ZnT8 scfv-CAR lentivirus was 2.4×108 TU/μL. The expression rate of GFP was 43.2% ± 4.1% in Tregs infected with ZnT8 scfv-CAR lentivirus. After being cultured in vitro for 14 days，the ZnT8-specific Tregs proliferated（634.3 ± 92.5）times，along with high expression of Foxp3（60.4% ± 3.5%）and Helios（64.3% ± 4.8%）. Conclusion：We obtained the PLVX-EGFP-ZnT8 scfv lentivector and synthesized ZnT8 scfv-CAR lentivirus successfully；Tregs infected with ZnT8 scfv-CAR lentivirus could maintain the CAR expression；CAR-Tregs were a group of CD4+CD25+ cells with high expression of Foxp3 and Helios.