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Studies on cell conversion methods by site-specific DNA demethylation

Research Project

Project/Area Number 24510284
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field System genome science
Research InstitutionNational Institute of Health Sciences

Principal Investigator

KUBOSAKI Atsutaka  国立医薬品食品衛生研究所, 衛生微生物部, 主任研究官 (30425673)

Research Collaborator TOMARU Yasunobu  
FURUHATA Erina  
SUZUKI Takahiro  
JAY Shin  
CHRISTOPHE Simon  
ANDO Yoshinori  
HASEGAWA Ryota  
HAYASHIZAKI Yoshihide  
SUZUKI Harukazu  
YOSHINARI Tomoya  
Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
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)
Keywordsエピゲノム制御 / ゲノム / 発現制御 / DNA脱メチル化
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.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • Research Products

    (3 results)

All 2013 2012

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (2 results)

  • [Journal Article] CpG site-specific alteration of hydroxymethylcytosine to methylcytosine beyond DNA replication2012

    • Author(s)
      Atsutaka Kubosaki
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 426 Issue: 1 Pages: 141-147

    • DOI

      10.1016/j.bbrc.2012.08.053

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] MoCEV; the modified cytosine monitoring system based on episomal vector during replication2013

    • Author(s)
      Harukazu Suzuki
    • Organizer
      Keystone Symposia
    • Place of Presentation
      New Mexico, USA
    • Related Report
      2012 Research-status Report
  • [Presentation] Modified cytosine monitoring system based on episomal vector during replication2012

    • Author(s)
      Atsutaka Kubosaki
    • Organizer
      MBSJ2012
    • Place of Presentation
      福岡
    • Related Report
      2012 Research-status Report

URL: 

Published: 2013-05-31   Modified: 2019-07-29  

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