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TChIP-Seq:细胞类型特异性表观基因组分析

TChIP-Seq: Cell-Type-Specific Epigenome Profiling.

作者信息

Mito Mari, Kadota Mitsutaka, Nakagawa Shinichi, Iwasaki Shintaro

机构信息

RNA Biology Laboratory, RIKEN; RNA Systems Biochemistry Laboratory, RIKEN Cluster for Pioneering Research.

Laboratory for Phyloinformatics, RIKEN Center for Biosystems Dynamics Research.

出版信息

J Vis Exp. 2019 Jan 23(143). doi: 10.3791/58298.

Abstract

Epigenetic regulation plays central roles in gene expression. Since histone modification was discovered in the 1960s, its physiological and pathological functions have been extensively studied. Indeed, the advent of next-generation deep sequencing and chromatin immunoprecipitation (ChIP) via specific histone modification antibodies has revolutionized our view of epigenetic regulation across the genome. Conversely, tissues typically consist of diverse cell types, and their complex mixture poses analytic challenges to investigating the epigenome in a particular cell type. To address the cell type-specific chromatin state in a genome-wide manner, we recently developed tandem chromatin immunoprecipitation sequencing (tChIP-Seq), which is based on the selective purification of chromatin by tagged core histone proteins from cell types of interest, followed by ChIP-Seq. The goal of this protocol is the introduction of best practices of tChIP-Seq. This technique provides a versatile tool for tissue-specific epigenome investigation in diverse histone modifications and model organisms.

摘要

表观遗传调控在基因表达中起着核心作用。自20世纪60年代发现组蛋白修饰以来,其生理和病理功能已得到广泛研究。事实上,下一代深度测序和通过特异性组蛋白修饰抗体进行的染色质免疫沉淀(ChIP)技术的出现,彻底改变了我们对全基因组表观遗传调控的看法。相反,组织通常由多种细胞类型组成,它们的复杂混合给研究特定细胞类型中的表观基因组带来了分析挑战。为了以全基因组方式研究细胞类型特异性染色质状态,我们最近开发了串联染色质免疫沉淀测序(tChIP-Seq),该技术基于通过标记的核心组蛋白蛋白从感兴趣的细胞类型中选择性纯化染色质,然后进行ChIP-Seq。本方案的目的是介绍tChIP-Seq的最佳实践。该技术为在多种组蛋白修饰和模式生物中进行组织特异性表观基因组研究提供了一种通用工具。

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