Synthetic Genomic Tools for Drug Discovery and Natural Products Functional Genomics (CLUSTER)
The goal of this demonstration project is to develop the capability to express complete biosynthetic pathways for natural products that are usually silent in their native microbial host (i.e. orphan biosynthetic gene clusters). Such a technology will increase our access to new chemical entities for potential use in drug development. By tailoring synthetic genomics techniques to the expression of biosynthetic gene clusters, we believe we will create the capacity to produce small molecules directly using synthetic expression constructs in platform production microorganisms. Publicly available high-throughput DNA sequencing data has already cataloged up to 20,000 orphan clusters; as a result, our approach has the potential to bring about the biological synthesis of a large number of natural products that are currently unavailable for evaluation as potential drugs. The proposed clones and expression constructs can be used as a starting point for evaluating the bioactivities of metabolites whose biosynthetic enzymes are encoded by a candidate orphan cluster. The techniques provided can subsequently be used as a blueprint for producing other new chemical entities from DNA sequence alone.
Two specific aims of this project will involve:
a) Re-engineering of Biosynthetic Gene Clusters: Demonstrate the capacity and expertise to rapidly and exhaustively re-engineer the transcriptional regulation of a characterized cluster and an orphan biosynthetic gene cluster.
b) Expression and analysis of re-engineered clusters: Demonstrate the capacity to transfer intact but completely re-engineered orphan clusters to an appropriate production host organism.
This project has been funded in whole or part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under contract numbers N01-AI30071 and/or HHSN272200900007C.