Gene Knockdown of Venezuelan Equine Encephalitis Virus E2 Glycoprotein Using DNA-Directed RNA Interference.

摘要

Venezuelan equine encephalitis virus is an important veterinary and human pathogen that also has the potential to be used as a bioterrorist agent. Since there are no approved vaccines or antiviral drugs for this virus, it is prudent that antiviral strategies be developed. RNA interference, an evolutionarily conserved but only recently discovered biological phenomenon, may be an effective gene manipulation tool to combat viruses. The ability of RNA interference to silence or knockdown specific mRNA through the use of short dsRNA fragments has been an effective tool to study gene function in many systems. In this study, we demonstrated the effectiveness of RNA interference to knockdown the VEE E2 gene expressed in mammalian cells. Here, a DNA-directed approach was used to transfect Vero cells with siRNA expression vectors. We demonstrated that both target siRNAs were effective in significantly reducing the level of E2 expression based on RT-PCR analysis of mRNA levels. Furthermore, the use of these vectors demonstrates the usefulness of a vector-based approach to silencing genes. Future studies will assess the efficacy of these E2- specific siRNA expression constructs in the inhibition of the VEE virus in vitro.