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Enzymatic synthesis of site specifically isotope labelled RNA

Research Project

Project/Area Number 15K13738
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Bio-related chemistry
Research InstitutionTokyo Institute of Technology

Principal Investigator

Seio Kohji  東京工業大学, 生命理工学院, 准教授 (20313356)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsヌクレオシド三リン酸 / RNAポリメラーゼ / アプタマー / 光ケージドRNA / ケージド核酸 / DNAポリメラーゼ / 同位体標識 / RNA / 光脱保護
Outline of Final Research Achievements

We studied the enzymatic incorporation of base- or sugar-modified ribonucleoside triphosphates by T7-RNA polymerase to develop the future technology to synthesize isotope-labeled RNA enzymatically. We tested 6-O-nitrobenzylguanosine and 4-O-nitrobenzyluridine triphosphates as the substrates of T7-RNA polymerase. However, it was revealed that both nucleoside triphosphates acted as inhibitor of the polymerase when concentrations of the nucleoside triphosphates were high. On the contrary, as the sugar modified nucleoside triphosphates, we tested 2'-O-carbamoyluridine triphosphate as the substrate or T7-RNA polyerase and found that this modified nucleoside triphosphate could be incorporated into RNA strand enzymatically. This result suggested that the carmaoyl-modified nucleoside triphosphate can be used for the enzymatic synthesis chemically modified functional RNAs such as RNA aptamers.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (10 results)

All 2017 2016 2015

All Journal Article (3 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 3 results,  Open Access: 3 results,  Acknowledgement Compliant: 2 results) Presentation (7 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Enzymatic synthesis and reverse transcription of RNAs incorporating 2 -O-carbamoyl uridine triphosphate2016

    • Author(s)
      Masaki Y, Ito H, Oda Y, Yamazaki K, Tago N, Ohno K, Ishii N, Tsunoda H, Kanamori T, Ohkubo A, Sekine M, Seio K.
    • Journal Title

      Chemical Communications

      Volume: 52 Issue: 87 Pages: 12889-12892

    • DOI

      10.1039/c6cc05796a

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] 7-(Benzofuran-2-yl)-7-deazadeoxyguanosine as a fluorescence turn-ON probe for single-strand DNA binding protein2016

    • Author(s)
      Munefumi Tokugawa, Yoshiaki Masaki, Jan Christian Canggadibrata, Kazuhei Kaneko, Takashi Shiozawa, Takashi Kanamori, Morten Grøtli, L. Marcus Wilhelmsson, Mitsuo Sekine and Kohji Seio
    • Journal Title

      Chemical Communications

      Volume: 52 Issue: 19 Pages: 3809-3812

    • DOI

      10.1039/c5cc09700b

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Photo-controlled binding of MutS to photo-caged DNA duplexes incorporating 4-O-(2-nitrobenzyl) or 4-O-[2-(2-nitrophenyl)propyl] thymidine2016

    • Author(s)
      Seio K, Ohno Y, Ohno K, Takeshita L, Kanamori T, Masaki Y, Sekine M.
    • Journal Title

      Bioorganic and Medicinal Chemistry Letters

      Volume: 26 Issue: 19 Pages: 4861-4863

    • DOI

      10.1016/j.bmcl.2016.07.075

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] ピリダジン-3-オンを核酸塩基部に有する核酸の核酸合成酵素に よる合成2017

    • Author(s)
      長岡健斗・友利貴人・竹下玲央・宮 武佑弥・正木慶昭・関根光雄・清尾康志
    • Organizer
      第97日本化学会春季年会
    • Place of Presentation
      慶応大学日吉キャンパス
    • Related Report
      2016 Annual Research Report
  • [Presentation] DNA ポリメラーゼによる取り込み能向上を目指した光ケージ ドヌクレオシド三リン酸の合成と性質2017

    • Author(s)
      竹下玲央・大野健太郎・長岡健斗・正木慶昭・清尾康志
    • Organizer
      第97日本化学会春季年会
    • Place of Presentation
      慶応大学日吉キャンパス
    • Related Report
      2016 Annual Research Report
  • [Presentation] DNA-タンパク質相互作用の光制御をめざした光ケージされたデオキシシュードウリジンを含む核酸の合成および性質2016

    • Author(s)
      竹下玲央 大野健太郎 正木慶昭 関根光雄 清尾康志
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学京田辺キャンパス
    • Year and Date
      2016-03-27
    • Related Report
      2015 Research-status Report
  • [Presentation] 塩基性条件下不安定なヌクレオシド三リン酸の新規合成法の開発2016

    • Author(s)
      小田雄貴 伊藤比佑吾 金森功吏 大窪 章寛 正木慶昭 関根光雄 清尾康志
    • Organizer
      日本化学会第96春季年会
    • Place of Presentation
      同志社大学京田辺キャンパス
    • Year and Date
      2016-03-26
    • Related Report
      2015 Research-status Report
  • [Presentation] Chemical and enzymatic synthesis of photo-caged DNA containing of N1 -nitrobenzyl-2'- deoxypseudouridine for UV dependent regulation of transcription2016

    • Author(s)
      Takeshita L, Ohno K, Masaki Y, Sekine M, Seio K.
    • Organizer
      The 43rd International Symposium on Nucleic Acids Chemistry
    • Place of Presentation
      熊本大学
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Synthesis and Enzymatic properties of C-nucleoside Triphosphate Containing Pyridazin-3- one as a uracil analog2016

    • Author(s)
      Tomori T, Nagaoka K, Miyatake Y, Masaki Y, Sekine M, Seio K.
    • Organizer
      The 43rd International Symposium on Nucleic Acids Chemistry
    • Place of Presentation
      熊本大学
    • Related Report
      2016 Annual Research Report
  • [Presentation] 光ケージヌクレオシド三リン酸の合成と性質2015

    • Author(s)
      大野健太郎 正木慶昭 金森功吏 関根光雄 清尾康志
    • Organizer
      第17回日本RNA学会年会
    • Place of Presentation
      ホテルライフォート札幌
    • Year and Date
      2015-07-15
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2018-03-22  

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