CONICET | Buscador de Institutos y Recursos Humanos

The alarming increase in antibiotic resistance has placed the focus on phages as an alternative antimicrobial therapy. Recently, the first patient treatment using engineered phages to combat a mycobacterial infection was successfully performed; genetic modifications were made using Bacteriophage Recombineering of Electroporated DNA (BRED). BRED is a simple technique that allows genetic manipulation of phages. The phage DNA and a recombination substrate, with short homology to the target, are co-electroporated into recombineering proficient bacteria promoting high levels of recombination. After electroporation, cells are recovered and plated in an infectious centre assay. Individual plaques are then screened by PCR to identify the mutant phage. The main characteristics of this technique, the advantages of engineered versus wild type phages for therapeutic purposes and the future perspective of BRED for doing such modifications, are reviewed here.