Application of Bacteriophage Genome Assembly Platforms

Using our knowledge of high-complexity Golden Gate Assembly we are applying bottom-up synthesis approaches to enable the rapid and flexible engineering of bacteriophage genomes. Engineered, synthetic bacteriophages have myriad applications in the study of phage biology, enabling unprecedented ease in establishing genotype-phenotype relationships. Further, phages offer the potential of developing new methods of bacterial genome manipulation, have potential as therapeutics for small-molecule resistant bacteria, and more. A bottom-up genome synthesis approach combines the targeted manipulation of genomes, everything from single nucleotide changes to the deletion/replacement of entire genes, with the ability to manipulate disparate regions of the genome simultaneously in a fast and highly efficient manner. This approach also eliminates the need for the sequential introduction of mutations and enables easy combinatorial screening through the mixing of WT of mutant parts in an assembly reaction. The flexibility of Golden Gate also allows assemblies to be quickly reconfigured to isolate specific regions of the phage genome for applications like library screening.

We have several collaborations with industry and academia to build phage engineering systems for a range of bacterial hosts and phage families.

 

image of a worklfow including Golden Gate DNA assembly, bacterial transformation and phage selection.

Sikkema, A.P., et al, (2023). Current Protocols. doi/10.1002/cpz1.882