Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy
Abstract
RNA interference (RNAi) is the process of sequence-specific post-transcriptional gene silencing triggered by double-stranded RNAs. In attempts to identify RNAi triggers that effectively function at lower concentrations, we found that synthetic RNA duplexes 25–30 nucleotides in length can be up to 100-fold more potent than corresponding conventional 21-mer small interfering RNAs (siRNAs). Some sites that are refractory to silencing by 21-mer siRNAs can be effectively targeted by 27-mer duplexes, with silencing lasting up to 10 d. Notably, the 27-mers do not induce interferon or activate protein kinase R (PKR). The enhanced potency of the longer duplexes is attributed to the fact that they are substrates of the Dicer endonuclease, directly linking the production of siRNAs to incorporation in the RNA-induced silencing complex. These results provide an alternative strategy for eliciting RNAi-mediated target cleavage using low concentrations of synthetic RNA as substrates for cellular Dicer-mediated cleavage.
Additional Information
© 2005 Nature Publishing Group. Received 22 July; accepted 1 November 2004; Published online 26 December 2004. D.K. is a Beckman Fellow. This work was supported by a grant from the Arnold and Mabel Beckman Foundation and the US National Institutes of Health (AI29329 and AI42552, and HL074704 to J.J.R.). The authors wish to dedicate this work to the memory of Arnold Beckman, who recently passed away.Attached Files
Supplemental Material - Fig1.pdf
Supplemental Material - Fig2.pdf
Supplemental Material - Fig3.pdf
Supplemental Material - Fig4.pdf
Supplemental Material - Table1.pdf
Supplemental Material - Table2.pdf
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Additional details
- Eprint ID
- 56299
- DOI
- 10.1038/nbt1051
- Resolver ID
- CaltechAUTHORS:20150401-144628864
- Arnold and Mabel Beckman Foundation
- AI29329
- NIH
- AI42552
- NIH
- HL074704
- NIH
- Created
-
2015-04-01Created from EPrint's datestamp field
- Updated
-
2021-11-10Created from EPrint's last_modified field