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Nature Biotechnology  23, 222 - 226 (2004)
Published online: 26 December 2004; | doi:10.1038/nbt1051

Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy

Dong-Ho Kim1, Mark A Behlke2, Scott D Rose2, Mi-Sook Chang3, Sangdun Choi3 & John J Rossi1, 4

1  Division of Molecular Biology, Beckman Research Institute of the City of Hope, 1450 East Duarte Rd., Duarte, California 91010, USA.

2  Integrated DNA Technologies, 1710 Commercial Park, Coralville, Iowa 52241, USA.

3  Division of Biology, 147-75, California Institute of Technology, Pasadena, California 91125, USA.

4  Graduate School of Biological Sciences, City of Hope and Beckman Research Institute of the City of Hope, 1450 East Duarte Rd., Duarte, California 91010, USA.

Correspondence should be addressed to John J Rossi jrossi@coh.org
RNA interference (RNAi) is the process of sequence-specific post-transcriptional gene silencing triggered by double-stranded RNAs1, 2, 3. 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.


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Nature Biotechnology
ISSN: 1087-0156
EISSN: 1546-1696
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