Construction and Characterization of Synthetic Dual-Input Promoters

Many genes are regulated at the level of transcriptional initiation by the interaction of multiple transcription factors within their promoter regions.  To shed light on the combined effects of multiple transcription factors in gene regulation, we used a synthetic and computational biology approach to construct and characterize dual-input gene regulatory networks in Saccharomyces cerevisiae. Inducible transcription factors, based on the GEV and Tet systems, were tested against a set of synthetic promoters with varying numbers of DNA binding sites. Expression of a GFP reporter in various concentrations of the inducers, estradiol and anhydrotetracycline, was used to quantify promoter activity and fit mathematical models. Understanding the effects of multiple transcriptional regulators will aid in the construction of predictable, large scale synthetic networks and provide a better understanding of natural gene regulatory networks.