Using gene-silencing linked to adaptation to find genes that increase antibiotic sensitivity and create a new plasmid selection marker

A CRISPR-based screening system identified the 'sensitasome,' a collection of genes that may sensitize bacteria to antibiotics.

• The screening system was applied to Escherichia coli and involved seven different antibiotics.
• Both known and previously unrecognized genes that influence antibiotic susceptibility were identified.
• Essential genes gyrA and gyrB were found to increase resistance to multiple antibiotics when repressed.
• Some genes conferred resistance to specific antibiotics while increasing susceptibility to others.
• The identified sensitasome-targeting spacers were effective in Shigella and Salmonella, suggesting a conserved resistance mechanism.
• A selected spacer was repurposed as a CRISPR-based antibiotic selection marker, offering a safer alternative to traditional methods.

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