Conversion of a Klebsiella pneumoniae Prophage towards a lytic Bacteriophage via Yeast-mediated Genome Editing

We investigated prophages from Klebsiella pneumoniae isolated from dogs and cats with urinary tract or respiratory infections. We isolated a functional prophage-derived myovirus, designated ?3. Employing a yeast-based synthetic genomics platform, we engineered ?3 derivative genomes harbouring deletions of either the gene coding for its integrase or its C2 lysogenic regulator, and rebooted them using E. coli as host. In vitro assays demonstrated enhanced lytic activity of rebooted phages against multi-drug-resistant K. pneumoniae strain 06KM907.