Practical guidelines for the comprehensive analysis of ChIP-seq data.

Practical guidelines for the comprehensive analysis of ChIP-seq data.
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DOI:
10.1371/journal.pcbi.1003326
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发表时间:
2013
影响因子:
4.3
通讯作者:
Zhang J
Zhang J
中科院分区:
生物学2区
文献类型:
--
作者:
Bailey T;Krajewski P;Ladunga I;Lefebvre C;Li Q;Liu T;Madrigal P;Taslim C;Zhang J

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绘制转录因子、核小体、组蛋白修饰、染色质重塑酶、分子伴侣和聚合酶的染色体位置是现代生物学的关键任务之一,如DNA元件百科全书(ENCODE)项目所证明的。为此,染色质免疫沉淀和高通量测序(ChIP-seq)是标准方法。使用ChIP-seq在体内绘制这种蛋白质-DNA相互作用不仅在样品制备和测序方面,而且在计算分析方面都面临着多重挑战。在这里,我们为ChIP-seq数据的计算分析提供了分步指南。我们解决了ChIP-seq数据分析中的所有主要步骤:测序深度选择,质量检查,映射,数据标准化,再现性评估,峰调用,差异结合分析,控制错误发现率,峰注释,可视化和基序分析。在指南的每一步,我们都会讨论一些最常用的软件工具。我们还强调了与ChIP-seq数据分析中每个步骤相关的挑战和问题。我们在图1中展示了一个用于分析ChIP-seq数据的简明工作流程,该流程补充并扩展了ENCODE和modENCODE项目的建议。工作流中的每个步骤将在以下部分中详细描述。
Mapping the chromosomal locations of transcription factors, nucleosomes, histone modifications, chromatin remodeling enzymes, chaperones, and polymerases is one of the key tasks of modern biology, as evidenced by the Encyclopedia of DNA Elements (ENCODE) Project. To this end, chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) is the standard methodology. Mapping such protein-DNA interactions in vivo using ChIP-seq presents multiple challenges not only in sample preparation and sequencing but also for computational analysis. Here, we present step-by-step guidelines for the computational analysis of ChIP-seq data. We address all the major steps in the analysis of ChIP-seq data: sequencing depth selection, quality checking, mapping, data normalization, assessment of reproducibility, peak calling, differential binding analysis, controlling the false discovery rate, peak annotation, visualization, and motif analysis. At each step in our guidelines we discuss some of the software tools most frequently used. We also highlight the challenges and problems associated with each step in ChIP-seq data analysis. We present a concise workflow for the analysis of ChIP-seq data in Figure 1 that complements and expands on the recommendations of the ENCODE and modENCODE projects. Each step in the workflow is described in detail in the following sections.
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