Harnessing CRISPR interference to resensitize laboratory strains and clinical isolates to last resort antibiotics

The global race against antimicrobial resistance requires novel antimicrobials that kill specific bacteria and minimize the emergence of new resistances. CRISPR/Cas-based antimicrobials have shown promising results in addressing killing specificity. However, the emergence of target sequence mutations triggered by Cas-cleavage poses a risk of generating new antibiotic-resistance gene (ARG) variants. A study evaluated an antibiotic re-sensitization strategy based on CRISPR interference (CRISPRi), which inhibits gene expression without damaging target DNA. The resistance to four antibiotics was significantly reduced by individual and multi-gene targeting of ARGs in recombinant E. coli. The study laid the foundation for further leveraging CRISPRi as an antimicrobial agent or research tool to selectively repress ARGs and investigate resistance mechanisms.