Mycobacterium tuberculosis remains a leading cause of infectious disease morbidity and mortality for which new drug combination therapies are needed. Combinations of respiratory inhibitors can have synergistic or synthetic lethal interactions suggesting that regimens with multiple bioenergetic inhibitors will drastically shorten treatment times. However, realizing this potential is hampered by a lack of on-target inhibitors and a poor understanding of which inhibitor combinations have the strongest interactions. To overcome these limitations, we have used CRISPR interference (CRISPRi) to characterize the consequences of transcriptionally inhibiting individual respiratory complexes and identify bioenergetic complexes that when simultaneously inhibited result in cell death. In this study, we identified known and novel synthetic lethal interactions and demonstrate how the engineering of CRISPRi-guide sequences can be used to further explore networks of interacting gene pairs. These results provide fundamental insights into the functions of and interactions between bioenergetic complexes and the utility of CRISPRi in designing drug combinations.

中文翻译：

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多重转录抑制确定了分枝杆菌呼吸复合物之间的杀菌相互作用网络

结核分枝杆菌仍然是需要新的药物联合疗法的传染病发病率和死亡率的主要原因。呼吸抑制剂的组合可能具有协同或合成致死相互作用，这表明具有多种生物能抑制剂的方案将大大缩短治疗时间。然而，由于缺乏靶向抑制剂以及对哪些抑制剂组合具有最强相互作用的了解不足，实现这一潜力受到阻碍。为了克服这些限制，我们使用 CRISPR 干扰 (CRISPRi) 来表征转录抑制单个呼吸复合物的后果，并确定同时抑制会导致细胞死亡的生物能复合物。在这项研究中，我们确定了已知的和新的合成致死相互作用，并展示了如何使用 CRISPRi 引导序列的工程来进一步探索相互作用的基因对网络。这些结果为生物能复合物的功能和相互作用以及 CRISPRi 在设计药物组合中的效用提供了基本见解。