Screening Target Genes Of Transcription Factor Ap-2 By Chromatin Immunoprecipitation Assay

In the post-genomic era,identifying and characterzing various DNA-protein interactions are a major challenge in the research of gene transcriptional regulation.Now,chromatin immunoprecipitation assay (CHIP) is ideally suited for studying DNA-protein interactions in vivo. Transcription factors tightly regulate gene expression in response to intra- and extracellular stimuli,and they often play a central role in determining cell fate by controlling the fundamental mechanism of gene transcription.Transcription factor AP-2 plays an important role in regulating transcription of a number of genes in mammalian development,differentiation and carcinogenesis.To identify the gene targets that mediate its effects,we optimized the condition of the chromatin immunoprecipitation assay and utilized the assay to screen unknown target genes of transcription factor AP-2 from human breast cancer cell line MDA-MB-453,cervical cancer cell line HeLa and mouse embryo.Cloning and sequencing of AP-2-ChIP-derived DNA fragments identified novel potential AP-2 target loci that were proximal to genes involved in the regulation of carcinogenesis,metabolism and others.Our results were consistent with the previous reports that ErbB2 was the target gene of AP-2.In this study,we utilized the assay to screen unknown target genes of transcription factor AP-2.Our results yielded a more complete understanding of AP-2-regulated signaling pathways and further elucidated the complex transcriptional networks.