Whole Genome Chromatin IP-Sequencing (ChIP-Seq) in Skeletal Muscle Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1668; p. 15
• Main Authors: So, Karl Kamhei, Peng, Xianlu Laura, Sun, Hao, Wang, Huating
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Animals; Binding Sites - genetics; Cell Line; Chromatin - genetics; Chromatin Immunoprecipitation - methods; Chromosome Mapping; Gene Library; Genome-Wide Association Study; Histones - genetics; Histones - metabolism; Mice; Muscle Fibers, Skeletal - metabolism; Transcription Factors - genetics; Transcription Factors - metabolism; Whole Genome Sequencing - methods; Transcription; ChIP-seq; Muscle cells
• ISSN: 1940-6029
• 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.