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Microscopic analysis of the morphological defects caused by CRISPRi essential gene depletion.

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posted on 2020-03-09, 17:39 authored by Robert C. Shields, Alejandro R. Walker, Natalie Maricic, Brinta Chakraborty, Simon A. M. Underhill, Robert A. Burne

(A) CRISPRi strains, by functional pathway, were grouped according to cell morphology phenotypes. Strains with depleted essential genes can have more than one morphological defect (e.g. increased cell chaining and heterogeneous sizes). (B) Representative micrographs for several of the cell morphology phenotypes that were observed are shown. Additional microscopy across all the pathways investigated can be found in S5–S12 Figs in S1 File. For sgRNA-atpA the arrow shows an area of increased nile red staining compared to other cells. The large round cells observed when silencing peptidoglycan biosynthesis is depicted with the sgRNA-murN micrograph. Arrows indicate potential anucleate cells when targeting the DNA replication-related gene dnaC.