Viral delivery of recombinant short hairpin RNAs.

Recent work demonstrates that RNA interference (RNAi) can coordinate protein expression. Inhibitory RNAs are expressed naturally in cells as microRNAs (miRNAs) or introduced into cells as small interfering RNAs (siRNAs). Both types of small RNAs can be used at the bench to silence mRNA expression. For many researchers, transfection of siRNAs synthesized in vitro or purchased from commercial sources is impractical for the cellular system under study. As an alternative to transfection-based methods, we provide a practical approach to accomplish siRNA-mediated gene silencing through the generation and introduction of recombinant viral vectors expressing short hairpin RNAs (shRNAs). shRNAs are subsequently processed to siRNAs in vivo, leading to efficient, and, in some cases, long-term silencing.