Dissecting The Regulatory Complexes Controlling Expression Of The Pluripotency Factors
Cellular identity is ultimately dictated by the interaction of transcription factors with regulatory elements (REs) to control gene expression. Genome-wide epigenome profiling techniques have significantly increased our understanding of cell-specific utilization of REs. However, it remains difficult to dissect the majority of factors interacting with these REs due to the lack of appropriate techniques. We developed TINC: TALE-mediated isolation of nuclear chromatin to dissect the factors responsible for cell-specific utilization of a RE. Using TINC we interrogated the protein complex formed at the Nanog promoter in embryonic stem cells, identifying known and novel complex members. To address the major limitations of single-locus proteomics technologies such as TINC, I’m currently optimizing tandem immunoprecipitation strategies to reduce non-specific enrichment of proteins.