Chinese Journal of Dermatology ›› 2011, Vol. 44 ›› Issue (11): 772-775.
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Abstract:
Objective To investigate the antiviral effect of siRNA on herpes simplex virus type 2 (HSV-2) in vitro. Methods Different siRNA duplexes targeting HSV-2 VP16 and DNA polymerase genes were designed and chemically synthesized. The antiviral effect of siRNA duplexes was evaluated by virus yield reduction assay. In detail, Vero cells cultured in 24-well plates were transfected with different siRNAs 4 hours before (preventive effect experiment) or 1 hour after (therapeutic effect experiment) the inoculation with HSV-2 (0.02 to 0.05 plaque forming unit/cell). After additional culture for 40 to 48 hours, the expression of target genes in Vero cells was measured by real-time quantitative reverse transcription (RT)-PCR. Results Both the siRNA duplex VP2-1 targeting HSV-2 VP16 and DP2-1 targeting DNA polymerase gene resulted in a remarkable decrease in viral titer by about 1 log10 in both the preventive and therapeutic effect experiment. As the preventive effect experiment showed, the expression of HSV-2 VP16 mRNA was decreased markedly by 58.6% and 79.5% by VP2-1, respectively, and that of HSV-2 DNA polymerase mRNA by 59.8%and 77.4% by DP2-1, respectively, at 6 and 12 hours after the infection. Conclusions The siRNA duplexes targeting HSV-2 VP16 and DNA polymerase genes could inhibit the proliferation of HSV-2. Hence, HSV-2 VP16 and DNA polymerase genes may serve as the target genes of RNA interference to inhibit HSV-2 replication.
Key words: siRNA
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