| Project/Area Number |
16H06692
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Biomolecular chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
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| Project Period (FY) |
2016-08-26 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 熱ゆらぎ / RNAアプタマー / 核酸ータンパク質相互作用 / 結合アフィニティー / 構造不均一性 / ハイスループット法 / RNA アプタマー / ハイスループットDNAシーケンシング / ハイスループットシーケンス / 熱揺らぎ / 核酸創薬 / 核酸 / 医薬 / ナノバイオ |
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| Outline of Final Research Achievements |
Aptamers are oligonucleotide ligands with specific binding affinity to target molecules. Here we develop a non-repeated, primer-less and target immobilization-free isolation method for generating RNA aptamers, which is robust to experimental noise. Uniquely, this method focuses on finding and removal of non-aptamer sequences from the RNA pool by RNase digestion leaving target-bound aptamer molecules, and thus is independent of aptamer types. The undigested RNA sequences remaining are so few in number that they must be mixed with a large excess of a known sequence for further manipulations and this sequence is then removed by restriction digestion followed by high-throughput sequencing analysis to identify aptamers. Using this method, we generated multiple RNA aptamers targeting thrombin and TGF-beta1 proteins, independently. This method also enables us to predict a common average structural property of these aptamers that is different from input RNA.
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