Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes.
In: eLife, 2020-11-06, S. 1-36
academicJournal
Zugriff:
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 be used to inform investigations of gene function. Exploiting this observation, we infer the existence of a mycobacterial restrictionmodification 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. [ABSTRACT FROM AUTHOR]
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Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes.
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Autor/in / Beteiligte Person: | de Wet, Timothy J. ; Winkler, Kristy R. ; Mhlanga, Musa ; Mizrahi, Valerie ; Warner, Digby F. |
Zeitschrift: | eLife, 2020-11-06, S. 1-36 |
Veröffentlichung: | 2020 |
Medientyp: | academicJournal |
ISSN: | 2050-084X (print) |
DOI: | 10.7554/eLife.60083 |
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