Read here: https://www.nature.com/articles/s41587-024-02392-z
In the new study, Fishman et al. introduce a phage genome engineering system that improves these methods. Their method can produce modifications at multiple sites in a phage genome rapidly, efficiently, and without the need for counterselection. To this end, the authors use retrons as an alternative source of donor DNA for homologous recombination. Retrons are bacterial retroelements that naturally defend bacteria against phage infection. These elements consist of a noncoding RNA and a reverse transcriptase. The reverse transcriptase uses a specific region in the noncoding RNA as a template to synthesize a single-stranded DNA. By replacing the noncoding RNA template with a sequence containing a desired edit, retrons can be used to produce donor DNA templates for genome engineering. To optimize retrons for phage genome modification, Fishman et al. combine the expression of a modified retron with a single-stranded annealing protein that ensures efficient annealing of the donor DNA to the lagging strand of the replication fork in the phage DNA…Â
Phage genome engineering with retrons
PHAGElogist 