| Project/Area Number |
16H04162
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Yoshimura Hideaki 東京大学, 大学院理学系研究科(理学部), 助教 (90464205)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2016: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
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| Keywords | RNA / 1分子計測(SMD) / ナノバイオ / イメージング / 細胞 / 蛍光プローブ / バイオイメージング / １分子計測(SMD) |
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| Outline of Final Research Achievements |
The target of this study is to develop a series of molecular probes to monitor RNA, from single molecule to single cell resolution, and from short to long observation time, using mPUM technology, which has bee originally established in our group. As an RNA probe that cover single-molecule and short-time observation, I developed a split-GFP based probe that addresses monitoring telomere repeat-containing RNA (TERRA), and observed TERRA, telomeres, and telomere-repeated protein in living cells simultaneously at the single molecule level. For visualization of RNA in single-cell resolution and long time domain, I developed split luciferase-based RNA probe that targets beta-actin mRNA in living cells. Using this probe, beta-actin mRNA in living cells were observed for tens of minutes at single cell resolution.
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| Academic Significance and Societal Importance of the Research Achievements |
近年様々な細胞生理機能について、RNAの動態や局在の直接関与が知られている。一方生細胞内RNAの動態・局在の可視化解析法は極めて限られていた。また、RNAは遺伝子発現の一次産物であり、生細胞内RNAの可視化定量法開発はすなわち非破壊的な遺伝子発現解析法の創出と言える。本研究では独自に構築したmPUMテクノロジーを用いて、1分子解像度での生細胞内RNA動態解析から、数10分間にわたる1細胞解像度RNA可視化定量法を構築した。すなわち本研究の成果により、生細胞内のRNA機能について1分子単位・秒単位の現象から細胞ごとに特徴が現れる長時間の遺伝子発現までを可視化解析する分析技術群の構築が実現した。
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