• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Pol III発現系を用いた哺乳動物細胞におけるRNA干渉の開発とがん治療への応用

Research Project

Project/Area Number 14030011
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionThe University of Tokyo

Principal Investigator

多比良 和誠  東京大学, 大学院・工学系研究科, 教授 (10261778)

Co-Investigator(Kenkyū-buntansha) 宮岸 真  東京大学, 大学院・工学系研究科, 特任助教授 (30323538)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥49,700,000 (Direct Cost: ¥49,700,000)
Fiscal Year 2004: ¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2003: ¥19,700,000 (Direct Cost: ¥19,700,000)
Fiscal Year 2002: ¥15,000,000 (Direct Cost: ¥15,000,000)
Keywordsジーンディスカバリー / アポトシス / RNA干渉 / RNAiライブラリー / RNA interference / pol III系 / U6プロモーター / 遺伝子治療 / がん遺伝子 / polIII系
Research Abstract

本研究課題では、siRNA発現ベクター系の最適化および、高機能化を行うと共に、現在作製段階であるヒト遺伝子に対するsiRNAベクターライブリーを用いて、がんに関連する新規機能遺伝子の探索、シグナリングパスウェイの解析を行っている。
今年度は、昨年度までに開発してきたsiRNA発現システムを用いて、アポトシスに関与する遺伝子数百個に対して、siRNA発現ベクターライブラリーを作製し、アポトシスシグナルトランスダクションの解析を試みた。タプシガルジン添加による小胞体(ER)ストレス依存的なアポトシスに対して、スクリーニングを行った結果、これまで知られていたDR5-Caspase8の経路に加えて、E2F1-PUMAパスウェイ、MAP2K8-ERKパスウェイが関与していること、また、ミトコンドリアの上流に、これまで報告のあるBid以外にも、PUMA、Badなど他の、BH3-only proteinが関与していることが新たに明らかになった。また、これまで関与していると考えられていたCaspase3、Caspase9非依存的な経路の存在が示唆された。今回の解析結果は、我々が開発したsiRNAベクターライブラリーを用いたスクリーニングがシグナル経路を解明する上で非常に効果的であることを強く示唆しており、今後のさらにスケールアップしたsiRNA発現ライブラリーを用いた、様々ながん細胞および、様々なアポトシス経路における新規機能遺伝子の網羅的探索を行っていく予定である。また、アポトシスを抑制するsiRNAベクターだけではなく、逆にアポトシスを促進するsiRNAベクターのスクリーングも、がんの治療に直接的に応用できる可能性があるので、精力的に行っていきたいと考えている。

Report

(3 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (19 results)

All 2004 Other

All Journal Article (6 results) Publications (13 results)

  • [Journal Article] Optimization of an siRNA-expression system with an improved hairpin and its significant suppressive effects in mammalian cells.2004

    • Author(s)
      Miyagishi, M., Sumimoto, H., Miyoshi, H., Kawakami, Y., Taira, K.
    • Journal Title

      Journal of Gene Medicine 6

      Pages: 715-723

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Know-how of RNA interference and its applications in research and therapy.2004

    • Author(s)
      Wadhwa, R., Kaul, S.C., Miyagishi, M., Taira, K.
    • Journal Title

      Mutat.Res. 567

      Pages: 71-84

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Use of a randomized hybrid ribozyme library for identification of genes involved in muscle differentiation.2004

    • Author(s)
      Wadhwa, R., Yaguchi, T., Kaur, K., Suyama, E., Kawasaki, H., Taira, K., Kaul, S.C.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 51622-51629

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification of metastasis-related genes using a library of randomized ribozymes in a mouse model.2004

    • Author(s)
      Suyama, E., Wadhwa, R., Kaur, K., Miyagishi, M., Kaul, S.C., Kawasaki, H., Taira, K.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 38083-38086

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification of a network involved in thapsigargin-induced apoptosis using a library of siRNA-expression vectors.2004

    • Author(s)
      Futami, T., Miyagishi, M., Taira, K.
    • Journal Title

      J.Biol.Chem. 280

      Pages: 826-831

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A20 is a negative regulator of interferon regulatory factor 3 signaling.2004

    • Author(s)
      Saitoh, T., Yamamoto, M., Miyagishi, M., Taira, K., Nakanishi, M., Fujita, T., kira, S., Yamamoto, N., Yamaoka, S.
    • Journal Title

      Journal of Immunology 174

      Pages: 1507-1512

    • Related Report
      2004 Annual Research Report
  • [Publications] Yoshinari, K, Miyagishi, M, Taira, K: "Effects on RNAi of structure, sequence and position within target RNA and potentially preferable sequence motif for siRNA activities in mammalian cells : Sequence itself rather than its location determines siRNA efficacy."Nucleic Acids Res.. 31. 691-666 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Taki, M., Kato, Y., Miyagishi, M., Takagi, Y., Taira, K.: "Small Interfering RNA Expression in Cells Based on an Efficiently Constructed Dumbbell-shaped DNA."Angew.Chem.Int.Ed.Engl.. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Miyagishi, M., Suminoto, H., Miyoshi, H., Kawakami, Y., Taira, K.: "Optimization of an siRNA-expression system with a mutated hairpin and its significant suppressive effects upon HIV vector-mediated transfer into mammalian cells."J.Gene Med.. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Miyagishi, M, Taira, K: "Strategies for generation of a stable siRNA-expression library directed against the human genome."Oligonucleotides. 13. 325-333 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yokota, T., Sakamoto, N., Enomoto, N., Tanabe, Y., Miyagishi, M., Maekawa, S., Yi, L, Kurosaki, M., Taira, K., Watanabe, M., Mizusawa, H.: "Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs."EMBO Rep.. 4. 602-628 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Onuki, R., Bando, Y., Suyama, E., Katayama, T., Kawasaki, H., Baba, T., Tohyama, M., Taira, K.: "Involvement of an RNA-dependent protein kinase(PKR) in tunicamycin-induced apoptosis and Alzheimer's disease."EMBO J.. 23. 959-968 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Miyagishi, M., Matsumoto, S., Futami, T., Akashi, H., Appasani, K., Takagi, Y., Sutou, S., Taira, K.: "In "RNAi technology", Appasani, K.(ed.), Cambridge University Press, Cambridge, UK."Generation of highly specific shRNAi expression libraries against entire human genome.(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Makoto Miyagishi: "U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targetes gene expression in mammalian cells"Nature Biotechnol.. 20・5. 497-500 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Makiko Hamada: "Effects on RNA interference in gene expression (RNAi) in cultured mammalian cells of mismatches and the introduction of chemical modifications at the 3' ends of siRNAs"Antisense Nucl. Acid Drug Develop.. 12・5. 301-309 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hiroaki Kawasaki: "Short hairpin type of dsRNAs that are controlled by tRNAVal promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cells"Nucleic Acids Res.. 31. 700-707 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takashi Futami: "Effects of an RNA helicase-recruiting sequence indirectly attached to antisense molecules"Antisense Nucl. Acid Drug Develop.. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hiroaki Kawasaki: "siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells"Nucleic Acids Res.. 31・3. (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Makoto Miyagishi: "Comparative analysis of suppressive effects of antisense, ribozyme and RNA interference in mammalian cells"Antisense Nucl. Acid Drug Develop.. (in press). (2003)

    • Related Report
      2002 Annual Research Report

URL: 

Published: 2002-04-01   Modified: 2018-03-28  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi