Development of technology to analyze dynamics of chromatin folding profiles at the single molecule level for understanding epigenome

2022 Fiscal Year Final Research Report

• Project/Area Number: 20H02591
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 28050:Nano/micro-systems-related
• Research Institution: The University of Tokyo
• Principal Investigator: Oana Hidehiro 東京大学, 大学院工学系研究科(工学部), 准教授 (20314172)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: マイクロ流体デバイス / 1細胞解析 / クロマチン / 染色体 / エピジェネティクス

Outline of Final Research Achievements

In this study, we developed a technique to isolate and unfold chromosomes from a single targeted mammalian-derived cell under a microscope in a microfluidic device, and fix the ends of the unfolded chromosomes to microstructures in microfluidic channels. This technique was applied to mouse-derived ES cells, and it was confirmed that chromosome unfolding does not occur uniformly along the chromatin, and that regions of easy unraveling and regions of difficult unraveling coexist. The partially unfolded chromosomes were then subjected to immunofluorescence staining for histone chemical modifications. The coincidence of one of the modifications accumulated in the transcription start region and the unfolded region of chromosome was visualized by single-molecule observation using a fluorescence microscope.

Free Research Field

バイオナノテクノロジー

Academic Significance and Societal Importance of the Research Achievements

本研究では、個々の細胞から染色体を取り出し、これを解きほぐして張力が均一な直線状の形態に保持する事を実現した。これにより、クロマチン折り畳みの動態やヒストン化学修飾の分布を精度良く光マッピングすることが可能となる。本技術を用いたクロマチン解析を更に進めて画像データ蓄積・解析する事によりエピジェネティクスについての理解が深まり、病気の診断や新たな知見に基づいた再生医療やがん治療法が生まれることが期待される。