Details

Autor(en) / Beteiligte

• de Wet, Timothy J
• Winkler, Kristy R
• Mhlanga, Musa
• Mizrahi, Valerie
• Warner, Digby F

Titel

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

Ist Teil von

• eLife, 2020-11, Vol.9

Ort / Verlag

England: eLife Sciences Publications, Ltd

Erscheinungsjahr

2020

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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, . 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 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.