Epigenetics in Human Health and Disease Laboratory, Baker IDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, Melbourne, Victoria 3004, Australia.
Biochimie. 2012 Nov;94(11):2353-9. doi: 10.1016/j.biochi.2012.06.002. Epub 2012 Jun 13.
The advent of massive parallel sequencing of immunopurified chromatin and its determinants has provided new avenues for researchers to map epigenome-wide changes and there is tremendous interest to uncover regulatory signatures to understand fundamental questions associated with chromatin structure and function. Indeed, the rapid development of large genome annotation projects has seen a resurgence in chromatin immunoprecipitation (ChIP) based protocols which are used to distinguish protein interactions coupled with large scale sequencing (Seq) to precisely map epigenome-wide interactions. Despite some of the great advances in our understanding of chromatin modifying complexes and their determinants, the development of ChIP-Seq technologies also pose specific demands on the integration of data for visualization, manipulation and analysis. In this article we discuss some of the considerations for experimental design planning, quality control, and bioinformatic analysis. The key aspects of post sequencing analysis are the identification of regions of interest, differentiation between biological conditions and the characterization of sequence differences for chromatin modifications. We provide an overview of best-practise approaches with background information and considerations of integrative analysis from ChIP-Seq experiments.
免疫纯化染色质及其决定因素的大规模平行测序的出现为研究人员提供了新的途径来绘制全基因组范围内的表观遗传变化图谱,并且人们非常有兴趣揭示调控特征,以了解与染色质结构和功能相关的基本问题。事实上,大型基因组注释项目的快速发展使得基于染色质免疫沉淀(ChIP)的方案重新兴起,这些方案用于区分与大规模测序(Seq)相结合的蛋白质相互作用,以精确绘制全基因组范围内的相互作用。尽管我们对染色质修饰复合物及其决定因素的理解取得了一些重大进展,但 ChIP-Seq 技术的发展也对数据的整合提出了可视化、操作和分析的具体要求。在本文中,我们讨论了实验设计规划、质量控制和生物信息学分析的一些考虑因素。测序后分析的关键方面是确定感兴趣的区域,区分生物学条件和染色质修饰的序列差异特征。我们提供了最佳实践方法的概述,包括背景信息和从 ChIP-Seq 实验进行综合分析的考虑因素。