Gene therapy involving RNA interference mediated by short hairpin RNAs (shRNAs) targeted against HIV-1 is a promising alternative to antiretroviral therapy.  Our lab has shown that fusing shRNA to carrier molecules, tRNAs, can provide a tunable level of shRNA delivery. We investigated how mutations in the acceptor stem of tRNA-serine affected release of fused shRNAs and whether variable cleavage of one tRNA-shRNA chimera impacted the knockdown efficiency of other shRNAs in the same construct. Acceptor stem mutations modulated the activity of fused shRNAs over a wide range. We observed only minor differences in knockdown efficiency by other shRNAs expressed in the same vector, demonstrating that the individual cassettes act largely independently. Strand selectivity was favorable for all constructs tested. Our results demonstrate that combined tRNA-shRNAs in a single vector do not significantly compete with one another and that efficient silencing can be achieved for multiple targets simultaneously with this system.

Effects of tRNA(Ser) acceptor stem mutations on competition amongst shRNAs

243rd National Meeting (2012)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.