Study of RNA Interference Induced Expression Silencing of DNA Repair Gene hMGMT

BACKGROUNDAIM: To construct the RNA interference vector which can specially induce the expression silencing of human DNA repair gene hMGMT. MATERIAL AND METHODS: The hMGMT specific siRNA expression cassette was made by two steps PCR,and then linked with pUC19to get pU6-MGMTi,co-transfected with pEGFP-C1into16HBE and screened by G418.The expression level of mRNA and protein was detected by RT-PCR and Western Blot,respectively. RESULTS: hMGMT-specific RNA interference vector pU6-MGMTi was constructed successfully.Transfected16HBE cells MGMT mRNA and protein expression level were dramatically suppressed. CONCLUSION: Silencing of MGMT expression cell line was built by RNA interference expression cassette and co-transfection technology,which offered condition for studying the gene function of MGMT.