Fully synthetic Golden Gate assembly system for engineering a Pseudomonas aeruginosa phiKMV-like phage (PNAS)

Sikkema and colleagues describe in PNAS an engineering platform for the phiKMV-like Pseudomonas aeruginosa phage 41S1 built on one-pot High Complexity Golden Gate assembly (HC-GGA): the genome is divided into synthesizable DNA fragments that assemble in a high-accuracy reaction, and phages are rescued by transformation into P. aeruginosa PAO1 or E. coli cells. Point mutations, insertions, deletions and fluorescence reporter genes were generated with near 100% success and minimal screening, providing a chassis for phages that broadly target P. aeruginosa.