Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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Outline of Final Research Achievements |
Hydroxymethylcytosine (hmC), one of several reported cytosine modifications, was recently thought to play an important role in DNA demethylation. To obtain the fundamental knowledge of hmC, methylcytosine dioxygenase was overexpressed in HEK293T cells and 5-hydroxymethyl deoxycytidines were analyzed by liquid chromatography-tandem mass spectrometry. I also used an original episomal vector-based method called MoCEV to monitor the fate of hmC beyond DNA replication in HEK293T cells. The MoCEV system containing fully hydroxymethylated-cytosine fragments revealed a significant modification towards methylcytosine after several rounds of DNA replication. Since the unmodified MoCEV did not undergo any DNA methylation during cell division, the results strongly suggest that somatic cells undergo hmC to methylcytosine specifically at the CpG sites during cell division. These data may help to induce site-specific DNA demethylation for cell conversion.
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