Model-based design of biological parts such as promoters remains an elusive goal of synthetic biology, especially for eukaryotes.  Promoters enable the expression of heterologous genes, provide regulation of genetic circuits, and establish the basis of metabolic engineering and the production of new biochemical products.   However, native promoters do not always allow for the desired expression level or regulation, which has created a demand for the generation of new promoters for a variety of cellular systems and applications.  While some successes have been noted - new promoters have been generated through random mutagenesis of native promoters, bioprospecting to find non-native promoters, and the creation of synthetic hybrid promoters - little progress has been made to rationally and systematically engineer promoters for a prescribed activity level.  Here, we successfully employ a method to both rationally change the expression level of native yeast promoters as well as design functional, purely synthetic yeast promoters using a model of nucleosome affinity.  In a single design cycle, this method was able to increase the expression of a native yeast promoter over three-fold, and create active synthetic promoters.  This work establishes the first DNA-level specification of promoter activity and is a first step toward the design and generation of promoters for any desired expression level and regulatory scheme.

Model-based design of synthetic yeast promoters via tuning of nucleosome architecture

247th National Meeting (2014)

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