Selective knockdown of Gs protein alphala subunit in primary cultured rat myocytes by RNA interference in vitro

Objective:To selectively knockdown the expression of a-subunit of the Gs protein (Gsα) in rat myocytes by RNA interference (RNAi),and to select the plasmids having the best effect on inhibiting Gsα.Method:Three pairs of shRNAs that target at gnas gene were designed,then the plasmids were transfected into primary cultured rat myocytes via liposome LipofectamineTM LTX PLUS,there are plasmids carrying a nonspecific shRNA coding sequence (HK plasmids) as the negative control group and the blank control group involved.The mRNA and protein expressions of Gsα were analyzed by semi-quantified RT-PCR and Western blot,respectively,and were normalized to internal control gene GAPDH.Result:The expressions of mRNA and protein of Gsα of test group were markedly decreased than that of control group (P0.01),while no significant difference was found in control group.Conclusion:RNAi can selectively knockdown Gsα expression in cultured myocytes,future studies are needed to explore the potential role of RNAi on Gsα function.