Studies on the induction methods of the DNA demethylation using next-generation sequencer

• Project/Area Number: 22710200
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: System genome science
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: KUBOSAKI Atsutaka 独立行政法人理化学研究所, LSA要素技術開発ユニット, 研究員 (30425673)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: メチル化DNA / 次世代シーケンサー / MBD-Seq / DNA脱メチル化誘導 / エピソーマルベクター / MBD-seq

Research Abstract

DNA methylation dynamically affects the expression of downstream genes. To obtain the fundamental technologies of site-specific changes of modified DNA, I first established the methylated DNA binding domain capture and next-generation sequencing(MBD-seq) and observed the DNA methylation status of human fibroblasts and monocytes. I also established a simple and flexible episomal vector-based method called MoCEV(Modified Cytosine in Episomal Vector) to monitor the status of cytosine modification of any desired DNA fragments. I observed enhanced expression of fluorescent marker protein controlled by DNA methylated promoter upon the treatment of 5-aza-2'-deoxycytidine in a dose dependent manner. I believe that the established methods and findings in this project may help to develop new methods of site-specific changes of modified DNA.

Research Products

• [Presentation] WP7 Activity : Experimental Validation by Epigenetic Markers (2011)
  • Author(s): Kubosaki, A., Simon, C., Suzuki, T., Shin, J., Hasegawa, Y., Suzuki, H.
  • Organizer: FANTOM5 Ume Blossom Meeting
  • Place of Presentation: Yokohama, Kanagawa, Japan