Synthesis of methylome based on basic operating principles of epigenetics

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K19276
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: Kyushu University
• Principal Investigator: Ito Takashi 九州大学, 医学研究院, 教授 (90201326)
• Project Period (FY): 2022-06-30 – 2024-03-31
• Keywords: DNAメチル化 / 構成的アプローチ / Cas9 / ナノポアシーケンシング

Outline of Final Research Achievements

Our challenge is to maintain artificially introduced methylated DNA regions using budding yeast as a model, which lacks endogenous DNA methylation. First, we established a genome editing method to introduce the entire E. coli λ phage DNA (~48.5 kb) into the HO locus as a target region to ensure that its methylation does not affect the physiological functions of budding yeast. At the same time, we improved the yeast genome editing vector series and established a system to detect genome editing and methylation using nanopore sequencing. In addition, to maintain exogenous DNA methylation, we constructed an inducible expression system for the maintenance DNA methylase from Cryptococcus neoformans and confirmed the expression and nuclear localization of the protein. These results laid the foundation for our investigation towards the ultimate goal

Free Research Field

ゲノム科学

Academic Significance and Societal Importance of the Research Achievements

外来性メチル化DNAの安定な維持が可能になると、エピジェネティック情報を保持した形でのクローニングが可能になり、様々な応用の可能性が拡がる。と同時に、メチル化領域の形成と維持も人工的に再現できれば、エピジェネティクス機構の構成的理解という意味でも大きな学術的意義が生じる。今回の成果は、こうした究極の目的に向けた挑戦の基盤が整備されたという点で一定の意義を有する。