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Short interfering RNA confers intracellular antiviral immunity in human cells

Nature volume 418, pages 430–434 (2002)Cite this article

A Corrigendum to this article was published on 08 May 2003

Abstract

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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Figure 1: Poliovirus growth suppressed by siRNA.
Figure 2: siRNA inhibits replication of poliovirus replicon in a sequence-dependent manner.
Figure 3: Gene silencing is rapid, post-transcriptional, and not dependent on viral replication.
Figure 4: Most cells treated with siC clear infection and do not harbour viral genomes.
Figure 5: Infected cells survive more than a day, but are eventually lysed by emerging siRNA-resistant virus.

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Acknowledgements

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.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, University of California, San Francisco, Box 0414, California, 94143-0414, USA

    Leonid Gitlin, Sveta Karelsky & Raul Andino

  2. Program in Neuroscience, University of California, San Francisco, California, 94143-0414, USA

    Leonid Gitlin

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  1. Leonid Gitlin
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Correspondence to Raul Andino.

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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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