ChIP on chip assays: genome-wide analysis of transcription factor binding and histone modifications.
Deregulation of transcriptional activity of many genes has been causatively linked to human diseases including cancer. Altered patterns of gene expression in normal and cancer cells are the result of inappropriate expression of transcription factors and chromatin-modifying proteins. Chromatin immunoprecipitation assay is a well-established tool for investigating the interactions between regulatory proteins and DNA at distinct stages of gene activation. ChIP coupled with DNA microarrays, known as ChIP on chip, allow us to determine the entire spectrum of in vivo DNA-binding sites for a given protein. This has been of immense value because ChIP on chip assays can provide a snapshot of the transcriptional regulatory mechanisms on a genome-wide scale. This article outlines the general strategies used to carry out ChIP-chip assays to study the differential recruitment of regulatory molecules based on the studies conducted in our lab as well as other published protocols.
Pillai
Smitha
S
Drug Discovery Program, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
Chellappan
Srikumar P
SP
D002457:Cell Extracts; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D016678:Genome / Q000235:genetics*; D006657:Histones / Q000378:metabolism*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011485:Protein Binding / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D014157:Transcription Factors / Q000378:metabolism*
D002457:Cell Extracts; D003432:Cross-Linking Reagents; D006657:Histones; D014157:Transcription Factors; D004247:DNA