Large scale CRISPRi-based whiB7 activation screen in Mycobacterium smegmatis
A genome-wide CRISPRi library (RLC11) was transformed into an M. smegmatis strain harboring the zeocin resistance gene driven by the whiB7 regulatory region. This library was plated in triplicate on plates containing either ATc only (control) or ATc + Zeocin (10 microgram/mL) to select for whiB7-expressing mutants. After 4 days of outgrowth, biomass was harvested from both sets of plates and the abundance of each CRISPRi knockdown strain was evaluated by deep sequencing the sgRNA-encoding region