Analysis of promoter-proximal pausing of transcription by using insertional chromatin immunoprecipitation(iChIP)

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To understand mechanisms of promoter-proximal pausing of transcription, we aimed to specifically isolate the promoter-proximal region of the c-fos gene and to identify molecules(proteins, DNA, RNA) binding to the isolated region by using insertional chromatin immunoprecipitation(iChIP). As a step of iChIP, we constructed a targeting vector for insertion of the binding elements of LexA, a bacterial DNA-binding protein, around the c-fos promoter-proximal region. We are now conducting gene targeting with the vector. On the other hand, we optimized iChIP for isolation of low copy number of a target genomic region and identification of proteins and RNA binding to the region.