Siobhan Cohen
Stony Brook University
Identifying Modulators of the Post-Antibiotic Effect in Pseudomonas aeruginosa Using CRISPRi
The post-antibiotic effect (PAE) is the persistent suppression of bacterial regrowth after the removal of an antimicrobial agent. Antibiotics that generate PAEs are dosed less frequently, improving patient adherence, reducing toxicity, and lowering treatment costs. PAEs can arise through several mechanisms, including antibiotic sequestration, reversible damage to cellular structures, and the extended occupancy of the drug target following drug elimination.
Previously, we demonstrated that the correlation between drug-target residence time (tR = 1/k off) and the post-antibiotic effect (PAE) can inform target vulnerability. CRISPR interference (CRISPRi) provides a complementary genetic method to assess target vulnerability by allowing tunable knockdown of specific essential proteins.
Using CRISPRi to transiently reduce target levels in Pseudomonas aeruginosa, we examine the growth defect as a function of time following washout of the inducer. This serves as a proxy measurement of the PAE, providing insight into the time-dependent inhibition of potential antibacterial targets.
The post-antibiotic effect (PAE) is the persistent suppression of bacterial regrowth after the removal of an antimicrobial agent. Antibiotics that generate PAEs are dosed less frequently, improving patient adherence, reducing toxicity, and lowering treatment costs. PAEs can arise through several mechanisms, including antibiotic sequestration, reversible damage to cellular structures, and the extended occupancy of the drug target following drug elimination.
Previously, we demonstrated that the correlation between drug-target residence time (tR = 1/k off) and the post-antibiotic effect (PAE) can inform target vulnerability. CRISPR interference (CRISPRi) provides a complementary genetic method to assess target vulnerability by allowing tunable knockdown of specific essential proteins.
Using CRISPRi to transiently reduce target levels in Pseudomonas aeruginosa, we examine the growth defect as a function of time following washout of the inducer. This serves as a proxy measurement of the PAE, providing insight into the time-dependent inhibition of potential antibacterial targets.
