Algorithmic DNA assembly methods for rapidly engineering multi-gene systems

Decreasing costs of DNA sequencing have revolutionized the life sciences by changing the types of questions we can ask about the living world and transforming the scale of investigations. As the cost to sequence a microbial genome de novo approaches $100, the availability of DNA sequence information in private and publically curated databases continues to explode. There are already tens of thousands of uncharacterized natural product biosynthetic gene clusters (BGCs) readily available in public sequence repositories. Synthetic DNA technologies are in the midst of a similar reduction in cost that we predict will revolutionize the life sciences once again. Our group is leveraging the latest DNA synthesis and assembly technologies to discover new natural products and engineer their biosynthesis. We have created an algorithmic DNA assembly pipeline that allows us to quickly build and screen libraries of several hundred permuted gene clusters. This pipeline was applied to optimize the genetics of a model refactored gene cluster. We are currently applying similar methods to allow the systematic investigation of natural product BGCs from soil bacteria.