Improvement of catalytic turnover based on transition state control of oligonucleotide template chemical reaction
| Project/Area Number |
16KT0052
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 特設分野 |
|---|
| Research Field |
Transition State Control
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
Abe Hiroshi 名古屋大学, 理学研究科, 教授 (80415067)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
木村 康明 名古屋大学, 理学研究科, 助教 (80769977)
|
|---|
| Project Period (FY) |
2016-07-19 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000)
Fiscal Year 2018: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2017: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2016: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
|
|---|
| Keywords | 核酸プローブ / 蛍光プローブ / 核酸鋳型反応 / 触媒回転数 / RNAプローブ / RNA / 鋳型反応 / DNA / 蛍光 / センサー / 遷移状態 / 触媒 / 増幅 |
|---|
| Outline of Final Research Achievements |
The fluorescence reaction based on the oligonucleotide (ON) template reaction is one of the most basic working principles of the RNA detection probe. In this method, an ON probe having a reactive functional group and an ON probe having a fluorescent precursor whose fluorescence is suppressed by a protecting group are associated on the target RNA, and then the protective group is removed and fluorescence signal is generated. Since this fluorescence reaction specifically occurs in the presence of target RNA, it can be the working principle RNA detection probe. In order to detect intracellular RNA highly sensitively, we worked on (1) accelerating ON template reaction and (2) development of a new method of intracellular administration of ON probe without lipofection. Prospective results were obtained in (1) by appropriately selecting the nucleophile for the reaction, and in (2) by use of phosphorothioate in the ON probe.
|
| Academic Significance and Societal Importance of the Research Achievements |
RNA検出法は、病気の診断、ウイルスの検出、細胞現象の解明など、様々な分野で重要な技術である。RNA検出では、いかに微量な標的RNAを検出できるか、すなわち検出感度の高感度化が重要な課題である。核酸検出プローブの作用原理である核酸鋳型反応の高速化により、高感度化が可能になるが、本研究ではその反応の高速化の指針を見出すことに成功した。また、核酸プローブの細胞投与において、既存の手法では望みでない蛍光シグナルを発してしまう問題があるが、本研究で開発した新たな投与法により、その問題を回避することが可能になった。両成果は、細胞内で微量なRNAの検出を可能にする技術の開発において重要な結果である。
|
Report
(3 results)
Research Products
(3 results)
