Gene silencing mediated by double-stranded RNA (dsRNA) is a sequence-specific, highly conserved mechanism in eukaryotes. In plants, it serves as an antiviral defence mechanism1,2,3. Animal cells also possess this machinery but its specific function is unclear4,5,6,7,8,9,10. Here we demonstrate that dsRNA can effectively protect human cells against infection by a rapidly replicating and highly cytolytic RNA virus. Pre-treatment of human and mouse cells with double-stranded, short interfering RNAs (siRNAs) to the poliovirus genome markedly reduces the titre of virus progeny and promotes clearance of the virus from most of the infected cells. The antiviral effect is sequence-specific and is not attributable to either classical antisense mechanisms or to interferon and the interferon response effectors protein kinase R (PKR) and RNaseL. Protection is the result of direct targeting of the viral genome by siRNA, as sequence analysis of escape virus (resistant to siRNAs) reveals one nucleotide substitution in the middle of the targeted sequence. Thus, siRNAs elicit specific intracellular antiviral resistance that may provide a therapeutic strategy against human viruses.
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Vance, V. & Vaucheret, H. RNA silencing in plants—defense and counterdefense. Science 292, 2277–2280 (2001)
Lindbo, J. A., Silva-Rosales, L., Proebsting, W. M. & Dougherty, W. G. Induction of a highly specific antiviral state in transgenic plants—implications for regulation of gene expression and virus resistance. Plant Cell 5, 1749–1759 (1993)
Voinnet, O., Pinto, Y. M. & Baulcombe, D. C. Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants. Proc. Natl Acad. Sci. USA 96, 14147–4152 (1999)
Bahramian, M. B. & Zarbl, H. Transcriptional and posttranscriptional silencing of rodent alpha1(I) collagen by a homologous transcriptionally self-silenced transgene. Mol. Cell Biol. 19, 274–283 (1999)
Dernburg, A. F., Zalevsky, J., Colaiacovo, M. P. & Villeneuve, A. M. Transgene-mediated cosuppression in the C. elegans germ line. Genes Dev. 14, 1578–1583 (2000)
Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811 (1998)
Kennerdell, J. R. & Carthew, R. W. Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway. Cell 95, 1017–1026 (1998)
Pal-Bhadra, M., Bhadra, U. & Birchler, J. A. Cosuppression in Drosophila: gene silencing of Alcohol dehydrogenase by white-Adh transgenes is Polycomb dependent. Cell 90, 479–490 (1997)
Svoboda, P., Stein, P., Hayashi, H. & Schultz, R. M. Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference. Development 127, 4147–4156 (2000)
Wianny, F. & Zernicka-Goetz, M. Specific interference with gene function by double-stranded RNA in early mouse development. Nature Cell Biol. 2, 70–75 (2000)
Bitko, V. & Barik, S. Phenotypic silencing of cytoplasmic genes using sequence-specific double-stranded short interfering RNA and its application in the reverse genetics of wild type negative-strand RNA viruses. BioMed Central Microbiol. 1, 34 (2001)
Caplen, N. J., Parrish, S., Imani, F., Fire, A. & Morgan, R. A. Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc. Natl Acad. Sci. USA 98, 9742–9747 (2001)
Elbashir, S. M. et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498 (2001)
Tolskaya, E. A. et al. Apoptosis-inducing and apoptosis-preventing functions of Poliovirus. J. Virol. 69, 1181–1189 (1995)
Kumar, M. & Carmichael, G. G. Antisense RNA function and fate of duplex RNA in cells of higher eukaryotes. Microbiol. Mol. Biol. Rev. 62, 1415–1434 (1998)
De Benedetti, A. & Baglioni, C. Inhibition of mRNA binding to ribosomes by localized activation of dsRNA-dependent protein kinase. Nature 311, 79–81 (1984)
Torrence, P. F. et al. Targeting RNA for degradation with a (2′-5′)oligoadenylate-antisense chimera. Proc. Natl Acad. Sci. USA 90, 1300–1304 (1993)
Olsen, P. H. & Ambros, V. The lin-4 regulatory RNA controls developmental timing in Caenorhabditis elegans by blocking LIN-14 protein synthesis after the initiation of translation. Dev. Biol. 216, 671–680 (1999)
Zamore, P. D., Tuschl, T., Sharp, P. A. & Bartel, D. P. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell 101, 25–33 (2000)
Baltimore, D. Gene therapy. Intracellular immunization. Nature 335, 395–396 (1988)
Ausubel, F. M. et al. (eds) Current Protocols in Molecular Biology (Green Publishing Associates and Wiley-Interscience, New York, 1994)
Zhou, A., Paranjape, J. M., Der, S. D., Williams, B. R. & Silverman, R. H. Interferon action in triply deficient mice reveals the existence of alternative antiviral pathways. Virology 258, 435–440 (1999)
Crotty, S. et al. Mucosal immunization of cynomolgus macaques with two serotypes of live poliovirus vectors expressing simian immunodeficiency virus antigens: stimulation of humoral, mucosal, and cellular immunity. J. Virol. 73, 9485–9495 (1999)
Herold, J. & Andino, R. Poliovirus requires a precise 5′ end for efficient positive-strand RNA synthesis. J. Virol. 74, 6394–6400 (2000)
Guidotti, L. G. et al. Viral clearance without destruction of infected cells during acute HIV infection. Science 284, 825–829 (1999)
Levine, B. et al. Antibody-mediated clearance of alphavirus infection from neurons. Science. 254, 856–860 (1991)
We are grateful to J. Frydman, L. Lanier, D. Ganem and A. Frankel for critical reading of the manuscript, and R. Silverman for providing the PKR- and RNaseL-deficient mouse embryonic fibroblasts. This work was supported by a grant from the National Institutes of Health to R.A.
The authors declare that they have no competing financial interests.
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Gitlin, L., Karelsky, S. & Andino, R. Short interfering RNA confers intracellular antiviral immunity in human cells. Nature 418, 430–434 (2002). https://doi.org/10.1038/nature00873
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