Development of DNA modification enzymes based on sequence-specificity of DNA binding domains and their application for genome editing.

2015 Fiscal Year Final Research Report

• Project/Area Number: 25410171
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Nomura Wataru 東京医科歯科大学, 生体材料工学研究所, 准教授 (80463909)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: タンパク質 / メチル化 / 人工酵素 / エピゲノム編集 / 核酸 / ケミカルバイオロジー / 合成生物学

Outline of Final Research Achievements

Improvement of genome editing tools such as CRISPR-Cas9 system leads to the increased demands for other gene modification tools such as DNA recombination and methylation. The split DNA methylase bearing split M.HhaI domains and zinc finger domains show the potential for the highly specific DNA methylation. Structural modelling study suggested that the DNA-split methylase complex could contain steric hindrance in some parts. Thus altered DNA assembly of split methylase on the target gene and several linker mutations were tested to obtain the increased activity in DNA methylation without losing sequence specificity. As the results, the assembly pattern in which ZFPs bind to the different strand on the target DNA showed increased activity in DNA methylation. In addition, the extension of linker length between the domains also affected on the increased activity. Above all, the results give new insights into the assembly of split protein domains especially on the target DNA.

Free Research Field

生物化学