Large scale chemical-genetic CRISPRi screen in Mycobacterium tuberculosis

Genome-wide CRISPRi libraries in M. tuberculosis H37Rv were grown in the presence of ATc to induce CRISPR interference at 1, 5, or 10 days prior to drug exposure. Drug screens were performed at concentrations spanning the predicted minimum inhibitory concentration (MIC). After 14 days outgrowth, genomic DNA were harvested from cultures treated with three descending doses of partially inhibitory drug concentrations (designated as "High", "Med", and "Low") , and analyzed sgRNA abundance by deep sequencing.