Institute of Basic Medical Sciences, Faculty of Medicine, Norwegian Center for Stem Cell Research, University of Oslo, Oslo, Norway.
Mol Biotechnol. 2010 May;45(1):87-100. doi: 10.1007/s12033-009-9239-8.
The biological significance of interactions of nuclear proteins with DNA in the context of gene expression, cell differentiation, or disease has immensely been enhanced by the advent of chromatin immunoprecipitation (ChIP). ChIP is a technique whereby a protein of interest is selectively immunoprecipitated from a chromatin preparation to determine the DNA sequences associated with it. ChIP has been widely used to map the localization of post-translationally modified histones, histone variants, transcription factors, or chromatin modifying enzymes on the genome or on a given locus. In spite of its power, ChIP has for a long time remained a cumbersome procedure requiring large numbers of cells. These limitations have sparked the development of modifications to shorten the procedure, simplify sample handling and make ChIP amenable to small numbers of cells. In addition, the combination of ChIP with DNA microarray and high-throughput sequencing technologies has in recent years enabled the profiling of histone modification, histone variants, and transcription factor occupancy on a genome-wide scale. This review highlights the variations on the theme of the ChIP assay, the various detection methods applied downstream of ChIP, and examples of their application.
染色质免疫沉淀（ChIP）技术的出现极大地增强了核蛋白与 DNA 相互作用在基因表达、细胞分化或疾病方面的生物学意义。ChIP 是一种从染色质制备物中选择性地免疫沉淀感兴趣的蛋白质，以确定与其相关的 DNA 序列的技术。ChIP 已广泛用于绘制组蛋白、组蛋白变体、转录因子或染色质修饰酶在基因组或特定基因座上的翻译后修饰的定位图谱。尽管 ChIP 具有强大的功能，但它长期以来一直是一项繁琐的程序，需要大量的细胞。这些限制激发了对缩短该程序、简化样品处理并使 ChIP 适用于少量细胞的改进的开发。此外，近年来，ChIP 与 DNA 微阵列和高通量测序技术的结合已能够在全基因组范围内对组蛋白修饰、组蛋白变体和转录因子占据进行分析。本文综述了 ChIP 检测方法的各种变化，以及下游应用的各种检测方法及其应用实例。
