Rna Interference Targeting Tnf Gene Treatment Of Rheumatoid Arthritis Experimental Research

OBSJECTIVETo construct a shRNA targeting TNF gene and to investigate the use of plasmid for transfection of siRNA into synovial cell of arthritis rheumatoid patients to achieve local RNA interference.METHODSTwe siRNA sequences of 19 nucleotides were derived from the full-length coding region of TNF gene. The shRNA templates were cloned into siRNA expression vector pGenesil-1 to generate TNF-targeting siRNA expression vector pGenesil-1 /TNF encoding 2 shRNAs targeting TNF by digestion and ligation step by step pGenesil-1/TNF was transfected into synovial cell, Real-time quantitive PCR was performed to validate the interfering efficacy of TNF at mRNA level.RESULTSAfter transfection the siRNA expression vector pGenesil-1 12hours and lasted 7days, the expression of TNF at RNA level were significantly down-regulated as validated by real-time quantitivc RT-PCR. Flow analysis revealed that RNA interference can successfully inhibit the expression of TNF in synovial cell.CONCLUSIONSRNA interference mediated by the siRNA expression down-regulate the vector pGenesil-1/TNF could expression of TNF, which lays a foundation for further research on RNA interference of rheumatoid arthritis.