Development of New methodology for Genome-wide Detection of Multi-contact Interaction between Chromatin Regions in Single Molecular Resolution

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21384
• 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: The University of Tokyo
• Principal Investigator: Fujiki Katsunori 東京大学, 定量生命科学研究所, 助教 (10646730)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: ゲノム / クロマチン高次構造 / 次世代シーケンサー / ロングリードシーケンサー / Hi-C / Pore-C / 多点間相互作用

Outline of Final Research Achievements

Through this project, I developed a new method, "targeted Pore-C," which enables the multi-contact analysis in genome-wide with a single molecular resolution. The original Pore-C method is a derivative version of the genome-wide contact detection method "Hi-C." It can detect multi-contact using a long-read sequencer and unfragmented library. I improved this Pore-C to focus on multi-contacts mediated by a specific factor of interest, which was achieved by introducing a chemical modification to DNA nearby the factor. Development of the new method in this project gained a sure success and remaining some challenges for practical use, such as improvement of modification efficiency.

Free Research Field

ゲノム生物学

Academic Significance and Societal Importance of the Research Achievements

クロマチン領域の相互の物理的位置関係はゲノム情報の発現に非常に重要な役割を果たしている。これらを解析する手法として現在はHi-CやHiChIPなどの方法が用いられてきたが、これら既存法は原理的にクロマチン高次構造を２領域間の点と点の相互作用の積算としてしか検出できない。今回Pore-C法をもとに開発したtargeted Pore-Cは、標的分子周辺に形成された多点間の相互作用を１分子の解像度で検出することを可能にし、近接領域を点と点ではなく塊で捉え、また細胞間のバリエーションを損なうことなく、既存技術では観察し得なかった核内のクロマチン高次構造を記述できる。