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siRNA-mediated gene silencing in vitro and in vivo

Nature Biotechnology volume 20, pages 1006–1010 (2002)Cite this article

Abstract

RNA interference is now established as an important biological strategy for gene silencing, but its application to mammalian cells has been limited by nonspecific inhibitory effects of long dsRNA on translation. Here, we describe a viral-mediated delivery mechanism that results in specific silencing of targeted genes through expression of small interfering RNA (siRNA). We establish proof of principle by markedly diminishing expression of exogenous and endogenous genes in vitro and in vivo in brain and liver, and further apply this strategy to a model system of a major class of neurodegenerative disorders, the polyglutamine diseases, to show reduced polyglutamine aggregation in cells. This viral-mediated strategy should prove generally useful in reducing expression of target genes to model biological processes or to provide therapy for dominant human diseases.

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Figure 1: siRNA expressed from CMV promoter constructs and in vitro effects.
Figure 2: Viral vectors expressing siRNA reduce expression from transgenic and endogenous alleles in vivo.
Figure 3: siGFP gene transfer reduces Q19-eGFP expression in cell lines.
Figure 4: siRNA-mediated reduction of expanded polyglutamine protein levels and intracellular aggregates.

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Acknowledgements

This work was supported in part by the NIH (HD 44093) and the Roy J. Carver Trust. The authors thank P.B. McCray, Jr. and M.J. Welsh for critical review of the work, and C. McLennan, J. Lindbloom, N. Kiewiet, and A. Espie-Ziemann for assistance.

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Author notes

  1. Haibin Xia and Qinwen Mao: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA

    Haibin Xia, Qinwen Mao & Beverly L Davidson

  2. Department of Neurology, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA

    Henry L Paulson & Beverly L Davidson

  3. Department of Physiology and Biophysics, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA

    Beverly L Davidson

Authors
  1. Haibin Xia
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Correspondence to Beverly L Davidson.

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Xia, H., Mao, Q., Paulson, H. et al. siRNA-mediated gene silencing in vitro and in vivo. Nat Biotechnol 20, 1006–1010 (2002). https://doi.org/10.1038/nbt739

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