New miRNA detecting system utilizing fluorescent DNA triplexes and rolling circle amplification

• Project/Area Number: 25620126
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: SEIO KOHJI 東京工業大学, 生命理工学研究科, 准教授 (20313356)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 蛍光核酸 / microRNA / 三重鎖核酸 / 遺伝子検出

Outline of Final Research Achievements

We planed to develop new microRNA detection techniques using fluorescent oligonucleotides and rolling circle amplification. As the fluorescent nucleosides developed 5-(3-methylbenzofuran-2-yl)deoxyuridine (dUMBF), the dUMBF derivatives incorporating various substituents on its benzofuran ring, and 7-deazaguanine incorporating benzofuranyl group at the 7 position. The fluorescent properties of these nucleosides and oligonucleotides incorporating them were studied.

Research Products

• [Journal Article] Controlling the fluorescence of benzofuran-modified uracil residues in oligonucleotides by triple-helix formation (2015)
  • Author(s): Takashi Kanamori, Hiroki Ohzeki, Yoshiaki Masaki, Akihiro Ohkubo, Mari Takahashi, Kengo Tsuda, Takuhiro Ito, Mikako Shirouzu, Kanako Kuwasako, Yutaka Muto, Mitsuo Sekine, Kohji Seio
  • Journal Title: Chembiochem Volume: 16 Issue: 1 Pages: 167-176
  • DOI: 10.1002/cbic.201402346
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Synthesis of Responsive Fluorescent Nucleobases 7-(Benzofuran-2-yl)-7-deazahypoxanthine and 7-(Benzofuran-2-yl)-7-deazaguanine Using Cross-coupling Reaction (2015)
  • Author(s): Munefumi Tokugawa, Kazuhei Kaneko, Masanori Saito, Takashi Kanamori, Yoshiaki Masaki, Akihiro Ohkubo, Mitsuo Sekine, Kohji Seio
  • Journal Title: Chemistry Letters Volume: 44 Issue: 1 Pages: 64-66
  • DOI: 10.1246/cl.140879
  • NAID: 130004699198
  • Peer Reviewed
• [Presentation] Synthesis and photophysical properties of 7-modified 7-deazaguanosine analogues (2014)
  • Author(s): Munefumi Tokugawa, Kazuhei Kaneko, Jan Cristian Canggadibrata, Takashi Kanamori, Yoshiaki Masaki, Akihiro Ohkubo, Marcus Wilhelmsson, Morten Grotli, Mitsuo Sekine, Kohji Seio
  • Organizer: The 41th international symposium on nucleic acids chemistry
  • Place of Presentation: Kitakyushu international conference hall
  • Year and Date: 2014-11-05 – 2014-11-07
• [Presentation] 3-メチルベンゾフラン誘導体を導入した新規蛍光ヌクレオチドの合成と蛍光特性
  • Author(s): 金森功吏 宮村大地 正木慶昭 大窪章寛 関根光雄 清尾 康志
  • Organizer: 日本化学会第９４春季年会
  • Place of Presentation: 名古屋大学 東山キャンパス
• [Presentation] ベンゾフラン環を導入した７－デアザヒポキサンチンの合成法とその蛍光特性
  • Author(s): 徳川宗史 金子和平 齋藤正憲 金森功吏 正木慶昭 大窪章寛 関根光雄 清尾康志
  • Organizer: 日本化学会第９４春季年会
  • Place of Presentation: 名古屋大学 東山キャンパス
• [Presentation] ７－（１－フェニルトリアゾル－４－イル）－７－デアザグアノシン誘導体の合成と性質
  • Author(s): 徳川宗史 金森功吏 正木慶昭 大窪章寛 関根光雄 WILHELMSSON MARCUS GROTLI MORTEN 清尾康志
  • Organizer: 日本化学会第９４春季年会
  • Place of Presentation: 名古屋大学 東山キャンパス
• [Presentation] 7-トリアゾリル-7-デアザグアニン誘導体の合成と蛍光特性
  • Author(s): 徳川宗史 Christopher Lawson, Anke Dierckx, 関根光雄 WILHELMSSON MARCUS GROTLI MORTEN 清尾康志
  • Organizer: 第７回バイオ関連化学シンポジウム
  • Place of Presentation: 名古屋大学豊田講堂