2006 Fiscal Year Final Research Report Summary
Development of the Intelligent Nucleic Acids having Multi Regulation Activity Triggered by Hybridization
| Project/Area Number |
17550158
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Tohoku University (2006)
Kyushu University (2005) |
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Principal Investigator |
NAGATSUGI Fumi Tohoku University, Institute of Multidiciplinary Research for Advanced Materials, Professor, 多元物質科学研究所, 教授 (90208025)
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| Project Period (FY) |
2005 – 2006
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| Keywords | Cross-Link / Hybridization / Drug releasing system / Fluorescence / Gene / FRET / Detection in Cells |
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| Research Abstract |
We have already established the strategy of synchronous activation by hybridization, in which the highly reactive cross-linking agent, 2-amino-6-vinylpurine nucleoside analog, can be generated from its stable precursors, the phenylsulfide derivatives, by a hybridization-promoted activation process with selectivity to cytosine. In this project, this in situ activation system was applied to method for the drug releasing system triggered by hybridization with the target sequence.
For the proof of our concept, we first designed the strategies for visualization of the releasing system in cells.
In 2004, we investigated that the fluorescent group was connected to 2-amino vinylpurine with sulfide bond and the release of sulfide group could be detected by increasing fluorescence but it did not work.
In 2005, we planed to exploit a fluorescence resonance energy transfer (FRET). We synthesized the 5'-fluorecein-labeled ODN containing 2-amino-6-vinylpurine at the position of 3 bases away from 5' end and 4-dimethylamino-azobenzene-thiol as a quencher have reacted to vinyl group. The fluorescence of double-labeled ODN probe decreased relative to the 5'-fluorecein single labeled ODN. When this probe hybridizes to exact complement and forms duplex DNA, the fluorescence increases gradually to be accompanied by release of the thiol group. These results have suggested that double-labeled ODN probe is capable of releasing the molecule triggered by hybridization in a highly sequence specific manner. The drug releasing system can be visualized by using our system in living cell.
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