Defect-complementation homologous recombination: A novel strategy for precise genome engineering of virulent phages.

ABSTRACT: Engineered bacteriophages (phages) have been developed to overcome the limitations of natural phage therapies and serve as precision-targeted agents against drug-resistant bacterial infections. However, their application has been constrained by the low efficiency of existing genome-editing tools, largely because of the absence of effective selection markers. This study proposed a novel strategy, termed defect-complementation homologous recombination (DCHR), for precise phage genome editing. In this approach, CRISPR-Cas9 cleaves a donor plasmid in host cells to release a linear donor template carrying homology arms, an essential phage gene used as a selection marker, and two lox sites. The donor template undergoes homologous recombination with the genome of essential gene-deficient p

SUBMITTER: Zhang H 

PROVIDER: S-EPMC12664982 | biostudies-literature | 2026 Jun

REPOSITORIES: biostudies-literature