Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes.

ABSTRACT: Mycobacterium tuberculosis possesses a large number of genes of unknown or predicted function, undermining fundamental understanding of pathogenicity and drug susceptibility. To address this challenge, we developed a high-throughput functional genomics approach combining inducible CRISPR-interference and image-based analyses of morphological features and sub-cellular chromosomal localizations in the related non-pathogen, M. smegmatis. Applying automated imaging and analysis to 263 essential gene knockdown mutants in an arrayed library, we derive robust, quantitative descriptions of bacillary morphologies consequent on gene silencing. Leveraging statistical-learning, we demonstrate that functionally related genes cluster by morphotypic similarity and that this information can

SUBMITTER: de Wet TJ 

PROVIDER: S-EPMC7647400 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature