Team:Heidelberg/Toolbox/Circularization

ABSTRACT

Promoters are the key regulators of gene expression. Possessing promoters which are active under a desired condition, at a desired strength and in a specified tissue is of great value for Plant Biotechnology, Gene Therapy and fundamental research in Bioscience. Therefore, it has become a desire to synthetically construct promoters responsive to a variety of pathways. We explore two ways to the synthesis of promoters: On one hand, we have developed a bioinformatical model and database (HEARTBEAT) describing the structure of promoters responsive to user-defined inputs. On the other hand, we have developed a biochemical method for the synthesis of randomized promoter libraries. Using this method, we have created a library of constitutive promoters of varying strength. Also, we have created libraries of promoters putatively responsive to a variety of pathways. We have screened these libraries for functional, pathway responsive promoters and present a detailed characterization of a NF-κB responsive promoter of our making. We finally discuss ways to combine randomized biochemical synthesis and bioinformatical modeling to propose a method towards the generation of promoters of complex regulation (i.e. by multiple pathways).