Abstract 1200 Expanding antimicrobial targets and new classes of growth inhibitors with CIMPLE-seq

Poor understanding of new antimicrobial mechanisms of action can impede compound prioritization for further development. Chemical genetics, which links bioactive compounds with their cellular targets, offers a streamlined approach to antibiotic discovery. Here, we generated a de novo chemical-genetic interaction dataset in the multi-resistant bacterium Burkholderia cenocepacia with a broad coverage of chemicals. Utilizing a CRISPR interference-mediated pooled library of essential genes (CIMPLE) followed by CRISPRi-seq (CIMPLE-seq), we screened 5,000 compounds, generating over 3.2 million chemical-genetic interactions.