? The objective of this proposal is to develop and commercialize an RNA polymerase II based transcription system reconstituted with highly purified recombinant and native transcription factors. This system will include three separated, but highly related, assay kits: 1) basal transcription kit that is able to detect basal level of transcription from most promoters for protein-encoding genes; 2) activator-dependent transcription kit that can be used to study transcriptional regulation by sequence-specific regulators, or to identify novel activators, co-activators, repressors, and new drugs that target individual transcription factors; 3) a reconstituted transcription kit that can be used to study transcriptional regulation on pre-assembled chromatin templates. The approaches will be based on the existing knowledge of the transcription machinery and the isolation of cDNAs encoding the general transcription factors, regulators, and chromatin remodeling factors. These studies would provide a broad spectrum of investigators, particularly those within the life science field, with a robust, well-developed and commercially available system to facilitate mechanistic analyses of transcription regulation and its linkage to human health. Our focus during Phase I period will be on expression and purification of general transcription factors and reconstitution of a basal RNA polymerase II based transcription system. ? ?
