Development of functional RNA whose activity of trapping a protein switches on/off for intra- and extracellular conditions

• Project/Area Number: 16K14678
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Kyoto University
• Principal Investigator: Katahira Masato 京都大学, エネルギー理工学研究所, 教授 (70211844)
• Research Collaborator: NAGATA Takashi 京都大学, エネルギー理工学研究所, 准教授 (10415250) ; YAMAOKI Yudai 京都大学, エネルギー理工学研究所, 博士研究員 (00778095)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: RNAアプタマー / Tat蛋白質 / スイッチング / NMR / 4重鎖 / カリウムイオン / Ｔａｔ蛋白質 / ＮＭＲ / ４重鎖 / 機能性核酸 / 構造変化 / FRET / 構造遷移 / 機能性RNA

Outline of Final Research Achievements

By utilizing an r(GGAGGAGGAGGA) molecule, whose structure drastically changes in response to K+, as a switching unit, we successfully developed the RNA aptamer whose activity of trapping the Tat protein switches on in response to K+. In response to the repeat of the increase and decrease of the K+ concentration, the activity repeatedly switches on and off. We also succeeded in observing NMR signals of nucleic acids in living human cells for the first time, which opened the way to explore the structures of nucleic acids in living human cells.

Research Products

• [Journal Article] The first successful observation of in-cell NMR signals of DNA and RNA in living human cells (2018)
  • Author(s): Yamaoki Yudai、Kiyoishi Ayaka、Miyake Masayuki、Kano Fumi、Murata Masayuki、Nagata Takashi、Katahira Masato
  • Journal Title: Phys Chem Chem Phys Volume: 20 Issue: 5 Pages: 2982-2985
  • DOI: 10.1039/c7cp05188c
  • NAID: 130007557810
  • Peer Reviewed / Open Access
• [Journal Article] Development of an RNA aptamer that acquires binding capacity against HIV-1 Tat protein via G-quadruplex formation in response to potassium ions (2017)
  • Author(s): Yamaoki, Y., Nagata, T., Mashima, T., Katahira, M.
  • Journal Title: Chemical communications Volume: 53 Issue: 52 Pages: 7056-7059
  • DOI: 10.1039/c7cc03312e
  • Peer Reviewed / Open Access
• [Presentation] Development of K+-responsive RNA aptamer and ribozyme, and in-cell NMR of nucleic acids in human cells (2017)
  • Author(s): Yamaoki, Y., Kiyoishi, A., Mashima, T., Kano, F., Murata, M., Nagata, T. and Katahira, M.
  • Organizer: International Symposium for NMR of Nucleic Acids
  • Place of Presentation: Ｇｙｅｏｎｓａｎｇ (韓国）
  • Year and Date: 2017-01-17
  • Int'l Joint Research
• [Presentation] Invention of RNA aptamer and ribozyme whose activi-ties switch on in response to potassium ion via G-quadruplex formation (2017)
  • Author(s): Y. Yamaoki, T. Mashima, T. Nagata, M. Katahira
  • Organizer: 6th International Meeting on Quadruplex Nucleic Acids
  • Int'l Joint Research
• [Presentation] In-cell NMR法によるヒト細胞内DNAおよびRNAの観測 (2017)
  • Author(s): 山置佑大，清石彩華，三宅雅之，加納ふみ，村田昌之，永田崇，片平正人
  • Organizer: 第56回NMR討論会
• [Presentation] nvention of K+-responsive Tat-binding RNA aptamer and hammerhead ribozyme, and in-cell NMR of nucleic acids (2016)
  • Author(s): Yamaoki, Y., Mashima, T., Nagata, T. and Katahira, M.
  • Organizer: The 43rd International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: 熊本大学（熊本）
  • Year and Date: 2016-09-27
  • Int'l Joint Research
• [Presentation] NMR studies of non-coding RNA and DNA recognition by TLS/FUS which induces transcriptional repression of cyclin D1 and telomere shortening (2016)
  • Author(s): Kondo, K., Mashima, T., Oyoshi, T., Yoneda, R., Kurokawa, R., Nagata, T. and Katahira, M.
  • Organizer: The XXVIIth International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: 京都国際会議場（京都）
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Remarks] 片平研究室
  • URL: http://www.iae.kyoto-u.ac.jp/bio/