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Activation of the interferon system by short-interfering RNAs

Nature Cell Biology volume 5, pages 834–839 (2003)Cite this article

Abstract

RNA interference (RNAi) is a powerful tool used to manipulate gene expression or determine gene function1,2. One technique of expressing the short double-stranded (ds) RNA intermediates required for interference in mammalian systems is the introduction of short-interfering (si) RNAs3,4,5,6. Although RNAi strategies are reliant on a high degree of specificity, little attention has been given to the potential non-specific effects that might be induced. Here, we found that transfection of siRNAs results in interferon (IFN)-mediated activation of the Jak–Stat pathway and global upregulation of IFN-stimulated genes. This effect is mediated by the dsRNA-dependent protein kinase, PKR, which is activated by 21-base-pair (bp) siRNAs and required for upregulation of IFN-β in response to siRNAs. In addition, we show by using cell lines deficient in specific components mediating IFN action that the RNAi mechanism itself is independent of the interferon system. Thus, siRNAs have broad and complicating effects beyond the selective silencing of target genes when introduced into cells. This is of critical importance, as siRNAs are currently being explored for their potential therapeutic use7,8.

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Figure 1: Intracellular effects of siRNA.
Figure 2: Global upregulation of ISGs in response to GAPDH siRNA in RCC1 cells.
Figure 3: PKR is necessary for the non-specific siRNA-induced signalling events.
Figure 4: RNA interference of GL3 luciferase expression by 21-bp GL3 siRNA.

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Acknowledgements

We would like to thank Z. Wang for synthesizing luciferase siRNAs, P. Stanhope-Baker for providing the Supplementary Information on the non-specific effects of the WT-1 siRNAs and A. Sadler for comments on the manuscript. This work was supported by National Institutes of Health (NIH) grants RO1 AI34039 and PO1 CA62220.

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Authors and Affiliations

  1. Department of Cancer Biology, Lerner Research Institute, NB40, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, 44195, OH, USA

    Carol A. Sledz, Michelle Holko, Michael J. de Veer, Robert H. Silverman & Bryan R.G. Williams

  2. Molecular Virology Program, Case Western Reserve University, Cleveland, 44106, OH, USA

    Carol A. Sledz, Robert H. Silverman & Bryan R.G. Williams

  3. Department of Genetics, Case Western Reserve University, Cleveland, 44106, OH, USA

    Michelle Holko & Bryan R.G. Williams

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  1. Carol A. Sledz
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  2. Michelle Holko
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  5. Bryan R.G. Williams
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Correspondence to Bryan R.G. Williams.

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Supplementary information

Supplementary Information, Fig. S1

Supplementary Information, Fig. S2 (PPT 1722 kb)

Supplementary Information, Fig. S3

Supplementary Information, Fig. S4

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Sledz, C., Holko, M., de Veer, M. et al. Activation of the interferon system by short-interfering RNAs. Nat Cell Biol 5, 834–839 (2003). https://doi.org/10.1038/ncb1038

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