Fig. 1: CRISPR screens reveal host factors required for TcsH intoxication.
From: Paeniclostridium sordellii hemorrhagic toxin targets TMPRSS2 to induce colonic epithelial lesions
a The sensitivities of several human cell lines, including HeLa, A549, U2OS, SHSY5Y, HepG2, 786-O, HT-29, Caco-2, and MCF-7, to TcsH were tested using the cytopathic cell-rounding assays. The percentages of the rounded cells were plotted over the TcsH concentrations. b Schematic of the screening process using TcsH on HT-29 Cas9 cells. GeCKOv2 and TKOv3 libraries were used independently. Genes enriched after 4 rounds of toxin selection from GeCKOv2 library (c) or TKOv3 library (d). The x-axis represents the number of gRNA reads for each targeted gene. The y-axis represents the number of unique gRNA. Top hints were marked with their gene names. e The sensitivities of the HT-29 TMPRSS2‒/‒, GMDS‒/‒, FUT4‒/‒, SLC35C1‒/‒, H2AFV‒/‒, UGT1A9‒/‒, GPC2‒/‒, and CNOT1‒/‒ cells to TcsH were measured using the cell-rounding assays. The percentages of the rounded cells were plotted over the TcsH concentrations. f The measured sensitivities in e are quantified and represented as CR50 (the toxin concentration leads to 50% cell rounding after 12–14 h toxin exposure) in a bar chart. In a, e, f, error bars (n = 6) indicate mean ± s.d.
