This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Development of a method to isolated and characterize native transcription factor complexes. The method utilizes triplex forming oligonucleotides to enrich native transcription factor complexes bound to chromatin in a sequence specific manner using magnetic beads. Triplex forming regions within DNA have long been described. This method involves: identification of such regions proximal to the regulatory regions of the gene/genes of interest;the enrichment of the native chromatin using biotinylated oligonucleotides which form a triple helix near the regulatory region;and the subsequent proteomic analysis of the intact transcription factor complexes attached to the enrich region of chromatin. A sequence specific ChIP experiment, so to speak.
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