Genome-wide CRISPRi-seq maps essential-gene contributions to aminoglycoside susceptibility in Pseudomonas aeruginosa

Genome-wide rhamnose-inducible CRISPRi in Pseudomonas aeruginosa PAO1 identifies 118 recurrent sensitizing and 120 recurrent protective gene knockdowns under aminoglycoside exposure.

• Why it matters: Essential genes could be antimicrobial targets, but how partial repression changes antibiotic susceptibility—and whether effects depend on condition—remains unclear.
• What they did: Built an 83,078-guide rhamnose-inducible CRISPRi library with mismatch guides for graded repression, then profiled fitness in Mueller-Hinton medium and fetal bovine serum, followed by aminoglycoside screens.
• The result: Individual CRISPRi knockdowns targeting atpI, accC, secA, and ftsZ enhanced tobramycin activity, and transcriptomes confirmed target silencing with polar effects and overlapping yet distinct responses.

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