Application of combined CRISPR screening for genetic and chemical-genetic interaction profiling in Mycobacterium tuberculosis

作者：Mei‐Yi Yan, Dandan Zheng, Si‐Shang Li, Xin-Yuan Ding, Chunliang Wang, Xiao-Peng Guo, Lingjun Zhan, Qi Jin, Jian Yang, Yicheng Sun · 发表于：Science Advances · 年份：2022 · DOI：10.1126/sciadv.add5907 · 被引用次数：27 · 研究领域：CRISPR and Genetic Engineering、Tuberculosis Research and Epidemiology、Mycobacterium research and diagnosis

CRISPR screening, including CRISPR interference (CRISPRi) and CRISPR-knockout (CRISPR-KO) screening, has become a powerful technology in the genetic screening of eukaryotes. In contrast with eukaryotes, CRISPR-KO screening has not yet been applied to functional genomics studies in bacteria. Here, we constructed genome-scale CRISPR-KO and also CRISPRi libraries in Mycobacterium tuberculosis (Mtb). We first examined these libraries to identify genes essential for Mtb viability. Subsequent screening identified dozens of genes associated with resistance/susceptibility to the antitubercular drug bedaquiline (BDQ). Genetic and chemical validation of the screening results suggested that it provided a valuable resource to investigate mechanisms of action underlying the effects of BDQ and to identify chemical-genetic synergies that can be used to optimize tuberculosis therapy. In summary, our results demonstrate the potential for efficient genome-wide CRISPR-KO screening in bacteria and establish a combined CRISPR screening approach for high-throughput investigation of genetic and chemical-genetic interactions in Mtb.