Functional system development towards screening of miRNA targets

• Project/Area Number: 19710171
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied genomics
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: KATO Yoshio National Institute of Advanced Industrial Science and Technology, セルエンジニアリング研究部門, 研究員 (20415657)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,780,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥480,000); Fiscal Year 2008: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2007: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: マイクロRNA / 標的遺伝子同定 / 遺伝子発現イメージング / 配列スクリーニング / フローサイトメトリー

Research Abstract

蛍光タンパク質遺伝子の非翻訳領域に、miRNA と完全に相補的な配列を挿入したウイルスベクターを構築し、miRNA の発現を生きた細胞内で検出する方法を構築した。筋肉特異的に発現するmiR-133 について、幹細胞から筋管細胞へと分化する際に発現上昇するmiR-133 の様子を生きた細胞内でレシオイメージングすることにより定量的に捉えることに成功した。本系はmiRNA の標的配列探索にも展開することが可能である。

Research Products

• [Journal Article] MicroRNA-140 is expressed in differentiated human articular chondrocytes and modulates IL-1 responses (2009)
  • Author(s): Shigeru Miyaki, Tomoyuki Nakasa, Shuhei Otsuki, Shawn P. Grogan, Reiji Higashiyama, Atsushi Inoue, Yoshio Kato, Tempei Sato, Martin K. Lotz, Hiroshi Asahara
  • Journal Title: Arth. Rheum. (in press)
  • Peer Reviewed
• [Journal Article] Real-time functional imaging for monitoring miR-133 during myogenic differentiation (2009)
  • Author(s): Yoshio Kato, Shigeru Miyaki, Shigetoshi Yokoyama, Shin Omori, Atsushi Inoue, Machiko Horiuchi and Hiroshi Asahara
  • Journal Title: Int. J. Biochem. Cell Biol. (in press)
  • Peer Reviewed
• [Journal Article] Detection of active microRNA species using fluorescent DNA probe. Miami Winter Symp (2008)
  • Author(s): Kittiphong Paiboonskuwong, Yoshio Kato
  • Journal Title: Short Rep. 19, T22
• [Journal Article] An efficient fluorescent mehod for selective detection of mature miRNA spiecies (2008)
  • Author(s): Yoshio Kato
  • Journal Title: Nucleic Acids Res. Symp. Ser. 52 Pages: 71-72
• [Journal Article] Detection of active microRNA species using fluorescent DNA probe (2008)
  • Author(s): Kittiphong Paiboonskuwong, Yoshio Kato
  • Journal Title: Proceedings of the Miami Winter Symposium 19
  • Peer Reviewed
• [Journal Article] HEKfectin トランスフェクション試薬を用いたHEK 細胞への遺伝子導入 (2007)
  • Author(s): 加藤義雄
  • Journal Title: BioRadiations 8 Pages: 40-41
• [Presentation] 継続的・定量的にmiRNA 発現を観測するための蛍光レポーターベクターの開発 (2009)
  • Author(s): 井上敦、味八木茂、加藤義雄
  • Organizer: 第4 回ケミカルバイオロジー学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2009-05-18
• [Presentation] An efficient fluorescent mehod for selective detection of mature miRNA spiecies (2008)
  • Author(s): Yoshio Kato
  • Organizer: Joint Symposium of 18th International Roundtable on Nucleosides, Nucleotides and Nucleic Acids (IRTXVIII) and 35th International Symposium on Nucleic Acids Chemistry (SNAC)
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2008-09-10
• [Presentation] Detection of active microRNA species using fluorescent DNA probe (2008)
  • Author(s): Kittiphong Paiboonskuwong, Yoshio Kato
  • Organizer: Miami Winter Symposium
  • Place of Presentation: Florida, USA
  • Year and Date: 2008-02-02
• [Presentation] Detection of active microRNA species using fluorescent DNA probe (2008)
  • Author(s): Yoshio Kato, Kittiphong Paiboonskuwong
  • Organizer: Miami Winter Symposium 2008
  • Place of Presentation: Miami,USA.
  • Year and Date: 2008-02-02
• [Presentation] 新規レポーターシステムによるmicroRNA の標的遺伝子同定" (2007)
  • Author(s): 井上敦、味八木茂、大森槙、浅原弘嗣、加藤義雄
  • Organizer: 遺伝情報DECODE 会議
  • Place of Presentation: 箱根
  • Year and Date: 2007-06-24
• [Presentation] 細胞内における低分子RNAの可視化に向けた蛍光DNA プローブの配列設計 (2007)
  • Author(s): 加藤義雄
  • Organizer: 第2 回ケミカルバイオロジー研究会
  • Place of Presentation: 京都
  • Year and Date: 2007-05-10