Chromatin Immunoprecipitation: Application to the Study of Asthma

Chromatin immunoprecipitation (ChIP) is a technique for studying interactions between proteins and DNA in living cells. A protein of interest is selectively immunoprecipitated from a chromatin preparation, to analyze the DNA sequences involved. ChIP can be used to determine whether a transcription factor interacts with a candidate target gene and to map the localization of histones with posttranslational modifications on the genome.The protein-DNA interactions are captured in vivo by chemical cross-linking. Cell lysis, DNA fragmentation, and immunoaffinity purification of the protein of interest allow to co-purify DNA fragments that are associated with that protein. The enriched protein-DNA population is ready to be quantified by PCR to detect precipitated DNA fragments. The combination of ChIP with DNA microarray analysis (ChIP-on-chip) and high-throughput sequencing (ChIP-seq) has enabled to obtain profiles of transcription factor occupancy sites and histone modifications throughout the genome.