Development of locus-specific single cell analysis of chromatin modifications in tissue

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K07372
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 48040:Medical biochemistry-related
• Research Institution: Yokohama City University
• Principal Investigator: NISHIYAMA Akira 横浜市立大学, 医学部, 准教授 (80589664)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: エピゲノム / ヒストン修飾 / トランスクリプトーム

Outline of Final Research Achievements

In this study, we established methods for analyzing gene expression, histone modifications, and chromatin structure at high resolution from very small number of cells. These methods are essential for analyzing transcriptional regulatory regions in the mouse tissues. For gene expression analysis, we performed RNA-seq for full length mRNA and total RNA. For histone modification analysis, we introduced methods for a smaller number of cells than our ChIP-seq method optimized for small amount of cells. In addition, we have combined the Hi-C method with capture probes to enable high-resolution chromatin structure analysis. Although we did not achieve the initial goal of establishing a method for single-cell epigenome analysis, these methods enable the high-precision analysis of transcriptional regulatory regions in mouse tissue.

Free Research Field

血液学

Academic Significance and Societal Importance of the Research Achievements

個体発生や臓器の形成はもちろんのこと、様々な疾患における病態理解においては、遺伝子発現がどのように調節されているか、またどのような異常が生じているかを研究することが非常に重要です。しかし、生体由来の微量サンプルでは遺伝子発現、ヒストン修飾などのエピゲエノム情報、高解像度のクロマチン高次構造の解析は困難でした。本研究では、生体由来の微量サンプルを用いて、高精細にこれらの解析を行う技術を確立しました。