Systematic and Comprehensive Analysis of Interferon Response induced by siRNA

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16300151
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomedical engineering/Biological material science
• Research Institution: The University of Tokyo
• Principal Investigator: MIYAGISHI Makoto The University of Tokyo, Faculty of Medicine, Project Associate Professor, 医学部附属病院, 特任助教授 (30323538)
• Project Period (FY): 2004 – 2005
• Keywords: Interferon Response / RNA interference / Biotechnology / siRNA

Research Abstract

In mammalian cells, siRNAs have been used to induce RNA interference (RNAi) in an attempt to prevent nonspecific effects (including the interferon (IFN) response) which are caused by long double-stranded RNAs (dsRNAs) of more than 30 bp. In this project, we developed a novel and simple strategy for avoiding activation of the IFN response by dsRNA, and analyzed interferon pathway by means of siRNA library. We showed that modified hairpin-RNAs (mhRNAs) of more than 100 bp, with multiple specific point-mutations within the sense strand and transcribed from the U6 or tRNAVa1 promoters, can cause RNAi without inducing the IFN pathway genes. These findings should enhance the exploitation of RNAi in mammalian cells, especially in the field of RNAi therapy against pathogenic viruses. Furthermore the results from screening by siRNA library showed that apoptosis pathway induced by dsRNA include a JNK/SAPK-mediated mitochondrial pathway and an ERK2-related pathway.

Research Products

• [Journal Article] Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype. (2006)
  • Author(s): Watanabe T, Sudoh M, Miyagishi, M., Akashi H, Arai M, Inoue K, Taira, K, Yoshiba M, Kohara M
  • Journal Title: Gene Ther. (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype. (2006)
  • Author(s): Watanabe T, Sudoh M, Miyagishi M, Akasi H, Arai M, Inoue K, Taira K, Yoshiba M, Kohara M.
  • Journal Title: Gene Ther. (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Escape from the interferon response associated with RNA interference using vectors that encode long modified hairpin-RNA (2005)
  • Author(s): Akashi, H., Miyagishi, M, Yokota, T., Watanabe, T., Hino, T., Nishina, K., Kohara, M., Taira, K.
  • Journal Title: Mol. BioSyst. 1 Pages: 382-389
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A20 is a negative regulator of IFN regulatory factor 3 signaling. (2005)
  • Author(s): Saitoh, T., Yamamoto, M., Miyagishi, M., Taira, K., Nakanishi, M., Fujuta, T., Akira, S., Yamamoto, N., Yamaoka, S.
  • Journal Title: Journal of Immunology 174 Pages: 1507-1512
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] siRNA becomes smart and intelligent. (2005)
  • Author(s): Miyagishi, M., Taira, K
  • Journal Title: Nature Biotech. 23 Pages: 946-947
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genome-wide application of RNAi to the discovery of potential drug targets. (2005)
  • Author(s): Ito, M, Kawano, M., Miyagishi, M., Taira, K.
  • Journal Title: Gene Ther. 12 Pages: 95-100
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Analysis of double-stranded RNA-induced apoptosis pathways using interferon-response noninducible small interfering RNA expression vector library. (2005)
  • Author(s): Matsumoto, S., Miyagishi, M., Akashi, H., Nagai, R., Taira, K.
  • Journal Title: J. Biol. Chem. 280 Pages: 25687-25696
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Gene therapy for human small-cell lung carcinoma by inactivation of Skp-2 with virally mediated RNA interference. (2005)
  • Author(s): Suminoto, H., Yamagata, S., Shimizu, A., Miyoshi, H., Mizuguchi, H., Hayakawa, T., Miyagishi, M., Taira, K., Kawakami, Y.
  • Journal Title: Gene Ther. 12 Pages: 95-100
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Escape from the interferon response associated with RNA interference using vectors that encode long modified hairpin-RNA (2005)
  • Author(s): Akashi, H., Miyagishi, M., Yokota, T., Watanabe, T., Hino, T., Nishina, K., Kohara, M., Taira, K.
  • Journal Title: Mol. BioSyst. 1 Pages: 382-389
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A20 is a negative regulator of IFN regulatory factor 3 signaling. (2005)
  • Author(s): Saitoh, T., Yamamoto, M., Miyagishi, M., Taira, K., Nakanishi, M., Fujita, T., Akira, S., Yamamoto, N., Yamaoka, S.
  • Journal Title: Journal of Immunology 174 Pages: 1507-1512
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] siRNA becomes smart and intelligent. (2005)
  • Author(s): Miyagishi, M., Taira, K.
  • Journal Title: Nature Biotech 23 Pages: 946-947
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genome-wide application of RNAi to the discovery of potential drug targets. (2005)
  • Author(s): Ito, M., Kawano, K., Miyagishi, M., Taira, K.
  • Journal Title: Gene Ther. 12 Pages: 95-100
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Analysis of double-stranded RNA-induced apoptosis pathways using interferon-response noninducible small interfering RNA expression vector library. (2005)
  • Author(s): Matsumoto, S., Miyagishi, M, Akashi, H., Nagai, R., Taira, K.
  • Journal Title: J. Biol. Chem. 280 Pages: 25687-25696
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene therapy for human small-cell lung carcinoma by inactivation of Skp-2 with virally mediated RNA interference. (2005)
  • Author(s): Sumimoto, H., Yamagata, S., Shimizu, A., Miyoshi, H., Mizuguchi, H., Hayakawa, T., Miyagishi, M., Taira, K, Kawakami, Y.
  • Journal Title: Gene Ther. 12 Pages: 95-100
  • Description: 「研究成果報告書概要(欧文)」より