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Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype

Gene Therapy volume 13, pages 883–892 (2006)Cite this article

Abstract

RNA interference (RNAi) can be used to inhibit viral replication in mammalian cells and therefore could be a powerful new antiviral therapy. Small interfering RNA (siRNA) may be effective for RNAi, but there are some technical problems that must be solved in each case, for example, predicting the effective siRNA target site and targeting heterogeneous sequences in a virus population. We show here that diced siRNA generated from long double-stranded RNA (dsRNA) is highly effective for inducing RNAi in HuH-7 cells harboring hepatitis C virus (HCV) replicons and can overcome variations in the HCV genotype. However, in mammalian cells, long dsRNA induced an interferon response and caused cell death. Here we describe an improvement of this method, U6 promoter-driven expression of long hairpin-RNA with multiple point mutations in the sense strand. This can efficiently silence HCV RNA replication and HCV protein expression without triggering the interferon response or cell death normally caused by dsRNA. In conclusion, intracellular-diced dsRNA efficiently induces RNAi, and, despite the high rate of mutation in HCV, it should be a feasible therapeutic strategy for silencing HCV RNA.

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Acknowledgements

We express their gratitude to Dr Christoph Seeger (Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA) for his kind gift of the replicon plasmid. We also thank Ms Etsuko Endo for creating the figures. This study was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan; the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation of Japan, and the Ministry of Health, Labor and Welfare of Japan.

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Authors and Affiliations

  1. Department of Microbiology and Cell Biology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan

    T Watanabe, M Arai, K Inoue & M Kohara

  2. Kamakura Research Labs, Chugai Pharmaceutical Co. Ltd, Kanagawa, Japan

    M Sudoh

  3. Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan

    M Miyagishi, H Akashi & K Taira

  4. Division of Gastroenterology, Showa University Fujigaoka Hospital, Yokohama, Japan

    T Watanabe, K Inoue & M Yoshiba

  5. Research & Development Division, Mitsubishi Pharma Corporation, Yokohama, Japan

    M Arai

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  1. T Watanabe
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Correspondence to M Kohara.

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Watanabe, T., Sudoh, M., Miyagishi, M. et al. Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype. Gene Ther 13, 883–892 (2006). https://doi.org/10.1038/sj.gt.3302734

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