| Project/Area Number |
15K13716
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Analytical chemistry
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| Research Institution | Tohoku University |
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Principal Investigator |
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| Project Period (FY) |
2015-04-01 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 小分子RNA / 2’-O-メチルRNA / 分離・検出 / アフィニティー電気泳動 / 分析科学 / 核酸 / 分子認識 |
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| Outline of Final Research Achievements |
Piwi-interacting RNA (piRNA), single stranded RNA of 26-31 nucleotide length, is a new class of small noncoding RNA which is mainly expressed in the germline. Their 3’-terminal nucleotides feature 2’-O-ribose methylation by 3’methyltransferase HEN1, which can enhance the piRNA stability. It has been known that piRNA plays a key role in the protection of germ cell genomes from the mobile genetic elements, thereby preventing transposon-induced defects in gametogenesis and fertility.
In this work, we have successfully improved boronate affinity electrophoresis for piRNA separation in the biological RNA samples, which is based on specific capture of boronic acid in the gel for cis-diol group in 3’terminal ribose of RNA species other than piRNAs having 2’-O-methylated ribose. Our improved method allows piRNA separation from mouse testes total RNA in a single step manner, revealing the great potential for quantitative separation and analysis of piRNA.
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