So Karl Kamhei, Peng Xianlu Laura, Sun Hao, Wang Huating
Department of Chemical Pathology, The Chinese University of Hong Kong, Hong Kong, China.
Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong, China.
Methods Mol Biol. 2017;1668:15-25. doi: 10.1007/978-1-4939-7283-8_2.
Transcriptional control of gene expression in skeletal muscle cell is involved in different processes ranging from muscle formation to regeneration. The identification of an increasing number of transcription factors, co-factors, and histone modifications has been greatly advanced by methods that allow studies of genome-wide chromatin-protein interactions. Chromatin immunoprecipitation with massively parallel DNA sequencing, or ChIP-seq, is a powerful tool for identifying binding sites of TFs/co-factors and histone modifications. The major steps of this technique involve immunoprecipitation of fragmented chromatin, followed by high-throughput sequencing to identify the protein bound regions genome-wide. Here, in this protocol, we will illustrate how the entire ChIP-seq is performed using global H3K27ac profiling in myoblast cells as an example.
骨骼肌细胞中基因表达的转录调控涉及从肌肉形成到再生的不同过程。允许对全基因组染色质-蛋白质相互作用进行研究的方法极大地推动了越来越多转录因子、辅助因子和组蛋白修饰的鉴定。染色质免疫沉淀结合大规模平行DNA测序（ChIP-seq）是鉴定转录因子/辅助因子结合位点和组蛋白修饰的强大工具。该技术的主要步骤包括对片段化染色质进行免疫沉淀，然后进行高通量测序以在全基因组范围内鉴定蛋白质结合区域。在此协议中，我们将以成肌细胞中全局H3K27ac分析为例说明整个ChIP-seq的操作过程。
