2011 Fiscal Year Final Research Report

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

Research Project

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
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
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.