Abstract: To investigate the effects of recombinant expression vector shRNA (small hairpin RNA) of HSV-2(Herpes simplex virus type 2) genes (including UL27 and UL54 genes) on HSV-2 replication, the recombinant expression vector shRNAs were transfected into HEK293 cells with liposome, and the transfection efficiency was detected by flow cytometry. The relative expression levels of UL27 and UL54 mRNA were detected by reverse transcription-real-time fluorescence quantitative PCR (RT-qPCR). The expression levels of UL27 and UL54 protein were detected by Western Blot, the titer of progeny virus was determined by TICD50, and the cell survival rate was detected by MTT method. The results showed that the inhibition rates of UL27 and UL54 mRNA expression in the group of UL27 shRNA75 + UL54 shRNA1081 were (88.10±1.95)% and (85.53±2.28)%, respectively. And the expression of gB protein and ICP27 protein and the titer of progeny virus were decreased significantly. The survival rate of HEK293 cells was (97.30±2.91)%. Compared with the single interference groups, the difference was statistically significant (P < 0.05). These results indicated that the combined interference of UL27shRNA75 and UL54shRNA1081 could significantly inhibit the replication of HSV-2 in HEK293 cells, which will provide a basis for the future research of HSV-2 multigene therapy.

Key words: human herpesvirus 2； small interfering RNA, UL27, UL54