Differential vulnerability CRISPRi screen in Rif Resistant rpoB H445Y and rpoB D435V mutant strains of M. tuberculosis
A genome-wide CRISPRi library (RLC0011) was transformed into an M. tuberculosis rpoB H445Y bioA deletion strain and an M. tuberculosis rpoB D435V bioA deletion strain The libraries were serially passaged in triplicate in either the presence (CRISPRi on) or absence (CRISPRi off) of ATc. Genomic DNA was harvested at the following generations (cumulative) and subjected to deep sequencing: 0, 2.5, 5, 7.5, 10, 15, 20, 25, and 30. Genetic vulnerability was then calculated, similar to what was described in Bosch &amp; DeJesus et al. (PMID: 34297925).