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.
Department of Biomedical and Diagnostic Sciences, University of Salamanca, Salamanca, Spain. chonela@usal.es.|Salamanca Institute for Biomedical Research (IBSAL), University Hospital of Salamanca, Salamanca, Spain. chonela@usal.es.
Asunción
A
García-Sánchez
Salamanca Institute for Biomedical Research (IBSAL), Salamanca, Spain.|Department of Clinical Biochemistry, University Hospital of Salamanca, Salamanca, Spain.
Fernándo
F
Marqués-García
D001249:Asthma / Q000235:genetics* / Q000378:metabolism; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D005786:Gene Expression Regulation; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D042421:Histone Code; D006657:Histones / Q000235:genetics*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011499:Protein Processing, Post-Translational
D006657:Histones
Antibody; ChIP-PCR; ChIP-on-chip; ChIP-seq; Chromatin; Cross-link; DNA; DNA-protein; Gene expression; Immunoprecipitation