| Project/Area Number |
16K05808
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | Saitama University |
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Principal Investigator |
SAITO Shingo 埼玉大学, 理工学研究科, 教授 (60343018)
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| Research Collaborator |
SUGANUMA Masami
YOSHIMOTO Keitaro
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | DNAアプタマー / 選抜 / キャピラリー電気泳動 / 細胞認識 / 細胞 / in vitro選抜 / 配列解析 / 電気泳動 |
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| Outline of Final Research Achievements |
A novel single-round DNA aptamer selection method for mammalian and microbial cells was successfully developed by means of a capillary electrophoresis (CE)-based methodology, called polymer-enhanced capillary transient isotachophoresis (PectI). The PectI separation yielded a single peak for the cells (particles) complexed with DNA aptamer candidates, which was effectively separated from a free randomized DNA library (molecules) peak. The cell peak including DNA aptamer sequences was fractionated by two-point detection CE system, then the fracion collected was served for next generation sequencing analysis. The sequences obtained by the PectI selection were subjected to binding assays, to prove those are cell-binding DNA aptamers.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究成果により細胞を認識するDNAアプタマーが既存法よりも簡便・高速に得られるようになった。特に本法は，解離反応速度の小さい高親和性アプタマーの獲得を指向した珍しい選抜法であり，既存法とは異なる種類のアプタマーの獲得が期待でき，この様に簡便に細胞を認識可能な分子を得ることによりDNAアプタマー創薬，臨床診断の分野だけでなく（細胞表面での）分子認識化学や細胞操作の学問分野を加速するためのキーテクノロジーとなりうると考えている。
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