Abstract

Mycobacterium tuberculosis remains a leading cause of infectious disease morbidity and mortality. Although a curable disease requiring a combination of four drugs for six months, drug-resistant strains require long treatment times (>18 months) with limited and often highly toxic drugs. New combination therapies that optimize the synergistic or synthetic lethal interactions between drugs have the potential to drastically reduce treatment times and increase cure rates. However, the identification of novel combination therapies is limited by available antibiotics that inhibit only a small number of biological targets. To address this lack in knowledge we have utilized a combination of mycobacterial CRISPR interference (CRISPRi) and phenotypic assays of bacterial growth and viability to identify respiratory complexes in Mycobacterium tuberculosis that when simultaneously inhibited result in cell death. The results from this study provide insight into the functional interactions between bioenergetic complexes, the clinical promise of targeting these processes and the general utility of CRISP Ri in designing optimised drug combinations for tuberculosis.