@article{DeWet2020,
abstract = {{\textless}i{\textgreater}Mycobacterium tuberculosis{\textless}/i{\textgreater} possesses a large number of genes of unknown or merely 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 molecular localizations in the related non-pathogen, {\textless}i{\textgreater}M. smegmatis{\textless}/i{\textgreater}. Applying automated imaging and analysis to an arrayed library of 272 essential gene knockdown mutants, 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 be used to infer gene function. Exploiting this observation, we reveal a previously unknown restriction-modification system, and identify filamentation as a defining mycobacterial response to histidine starvation. Our results support the application of large-scale image-based analyses for mycobacterial functional genomics, simultaneously establishing the utility of this approach for drug mechanism-of-action studies.},
author = {de Wet, Timothy J and Winkler, Kirstin and Mhlanga, Musa and Mizrahi, Valerie and Warner, Digby F},
doi = {10.1101/2020.03.20.000372},
file = {:C$\backslash$:/Users/01462563/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/de Wet et al. - 2020 - Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes.pdf:pdf},
journal = {bioRxiv},
keywords = {OA,fund{\_}ack,original},
mendeley-tags = {OA,fund{\_}ack,original},
month = {mar},
pages = {2020.03.20.000372},
publisher = {Cold Spring Harbor Laboratory},
title = {{Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes}},
url = {https://doi.org/10.1101/2020.03.20.000372},
year = {2020}
}

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Applying automated imaging and analysis to an arrayed library of 272 essential gene knockdown mutants, 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 be used to infer gene function. Exploiting this observation, we reveal a previously unknown restriction-modification system, and identify filamentation as a defining mycobacterial response to histidine starvation. 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