2019 Fiscal Year Final Research Report
Development of functional nucleic acid whose activity is altered between inside and outside the cell by utilizing in-cell NMR
Project/Area Number |
17K14515
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Research Category |
Grant-in-Aid for Young Scientists (B)
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Allocation Type | Multi-year Fund |
Research Field |
Bio-related chemistry
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Research Institution | Kyoto University |
Principal Investigator |
YAMAOKI Yudai 京都大学, エネルギー理工学研究所, 研究員 (00778095)
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Project Period (FY) |
2017-04-01 – 2020-03-31
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Keywords | in-cell NMR / 四重鎖核酸 / リボザイム / アプタマー / カリウムイオン |
Outline of Final Research Achievements |
The potassium ion concentration inside cells (100-150 mM) is much higher than that outside the cells (3.5-5.5 mM), therefore functional RNAs that sense the potassium ion concentration can switch their activities ON/OFF between the intra- and extracellular environments. Here we developed an RNA aptamer whose binding activity is altered in response to potassium ion via quadruplex structure formation. Furthermore, in order to monitoring the functional RNA inside the living cells, we tried to measure in-cell NMR spectra of RNAs in living human cells. Here NMR signals of DNA and RNA were successfully observed in living human cells. The observed signals indicated the formation of DNA and RNA hairpin structures in living human cells.
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Free Research Field |
構造生物学
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Academic Significance and Societal Importance of the Research Achievements |
カリウムイオン濃度を感知して細胞内に取り込まれた時のみ活性がオンになる新規機能性RNA分子は細胞内分子を標的とする核酸医薬品および同分野で課題となっているドラッグデリバリーシステム開発の一助と成り得る。また、細胞内は試験管内環境とは異なり、様々な生体分子が密に詰まった分子混雑環境である。このような混雑環境中では核酸の挙動が試験管内環境とは異なる。本研究で確立したヒト細胞を用いた核酸のin-cell NMR法はこのような分子混雑環境下で核酸の挙動を詳細に調べる手法と成り得る。これは核酸の制御する様々な生命現象を調べるツールと成ることが期待される。
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