Study On The Expression Of Bak Gene On Dc Inhibited By Special Sirna

The effect of expression of proapoptotic gene BAK on human dendritic cells (DCs) were investigated with specific small interfering RNA in this study.Two highly specific siRNA candidates targeting BAK gene were designed, and their recording sequences were constructed into the recombinant plasmids psi-BAK-183 and psi-BAK-195. After validation through DNA restriction enzyme digestion and specific DNA fragment sequencing, both recombinant plamids and control plasmid were transfected into DCs with liposome. After transfection for 72 hours, the expression of BAK gene in DCs were detected with Western-blot, and the viablel rate of DCs were detected with flow cytomertry.The results of Western Blot showed that two special siRNA could inhibite the expression of Bak up to (51.5±3.54)﹪and (26.5±6.36)﹪respectively at 72 hours after transfection;the viable rate of DC at 72 hours after transfection with recombinant plasmids were 93.95﹪and 89.66﹪, both which higher than 88.00﹪of control.si-BAK-183,si-BAK-195 can reduce the expression of proapoptotic gene BAK in human dendritic cell. This work supplied an experiment base for prolonging the life of dendritic cells,meanwhile, provided a new strategy and approach for enhancing antigen presenting capability.