Objective To establish a HEK-293T cell model with sodium channel function similar to that in PRKAG2 syndrome. Methods Vectors of PRKAG2 gene and SCN5A gene were constructed and were used to co-transfect HEK-293T cells. Immunofluorescence, real-time PCR and Western blotting analysis were used to examine the transfection results and the expression of interested mRNA and protein. Results Imunofluorescence findings showed red fluorescence and green fluorescence in HEK-293T cells 48 hours after co-transfection. Real-time PCR and Western blotting analysis showed SCN5A expression in pure SCN5A group, SCN5A + wild-type group, SCN5A + R302Q mutant group, and SCN5A + G100S mutant group, but not in the blank control group, showing significant significance(P<0.05). Expression of PRKAG2 gene and protein in SCN5A+PRKAG2 group was significantly higher than those in the blank control group, pure SCN5A group, SCN5A+R302Q, and SCN5A+G100S groups (P<0.05) . Conclusion We have successfully established a HEK-293T cell model with sodium channel function similar to that in PRKAG2 syndrome, paving a way for future study.