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Alleviation of off-target effects from vector-encoded shRNAs via codelivered RNA decoys
- Mockenhaupt, Stefan, Grosse, Stefanie, Rupp, Daniel, Bartenschlager, Ralf, Grimm, Dirk
- Proceedings of the National Academy of Sciences of the United States of America 2015 v.112 no.30 pp. E4007
- Hepatitis C virus, RNA, RNA interference, biomedical research, gene expression, gene therapy
- Induction of RNAi through shRNA expression holds great potential for biomedical research. Ideal shRNAs exhibit high specificity, potency, and safety, but options for rationally designing such shRNAs remain limited. One ensuing concern is unintended perturbation of gene expression by shRNA sense or antisense strands (off-targeting). Here, we report a novel solution that specifically counteracts adverse activity of shRNA sense strands via their sequestration by tough decoy RNAs (TuDs). Using hepatitis C virus as a clinically relevant example, we demonstrate how TuD coexpression from an Adeno-associated viral (AAV) gene therapy vector concurrently decreases shRNA off-targeting and enhances on-target inhibition. The versatility of TuDs and AAVs makes our new strategy useful for improving specificity and safety of a wide array of RNAi experiments.