| Project/Area Number |
16K07224
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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 |
System genome science
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Aoki Hiroyoshi 国立研究開発法人理化学研究所, 光量子工学研究センター, 研究員 (50518636)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2018: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2017: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | １細胞ゲノム解析 / ハイドロゲル・マイクロカプセル / 難培養微生物 / アガロースゲル・マイクロカプセル / 全ゲノム増幅 / 一菌体ゲノム解析 / ハイドロゲルマイクロカプセル / 増幅バイアス / 全ゲノム酵素増幅 / シングルセル・ゲノミクス / マイクロリアクタ |
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| Outline of Final Research Achievements |
Single cell genomics (SCG) analyse genetic information of single cell from trace amount of sample. The SCG is expected to innovate biology, microbiology, and medicine. However, infinitesimal genomic DNA from single cell caused uneven enzymatic DNA amplification for DNA sequencing, which prevented to analyse all genes on the genome. To improve the uniformity of the DNA amplification, agarose gel microcapsule (AGM) is developed. AGM consists of agarose gel shell and alginate sol core with easily preparing using commercially available inexpensive reagents, plasticware, and laboratory equipment. Pico-litre-scale DNA amplification in the AGM core drastically improved the DNA amplification uniformities over 90% compared to conventional method (30%). Thus, AGM will accelerate the SCG especially for analyses of environmental microbiome, early diagnosis of cancer metastasis, and quality control of regenerative medicine.
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| Academic Significance and Societal Importance of the Research Achievements |
１細胞染色体DNAの酵素増幅では，マイクロ流路やエマルジョン（オイル中に懸濁した微小液滴）による均一化は報告されていたが，特殊な機器が必要であり，壊れやすいエマルジョンの単離が困難，などの課題があった．本法は多くの研究者がより簡便に１細胞ゲノム解析を行えるよう，入手しやすい試薬と実験器具で作製でき，安定に単離できるAGMを開発した．学術的には半透過性のAGM内の酵素増幅による，増幅量の増加と均一性向上を示し，社会的には１細胞染色体DNAの酵素増幅の均一性の向上により，高品質の遺伝子情報を提供する手段を提供し，生物，微生物，および医学の発展に寄与した．
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