Fig. 1: Genome-wide CRISPR–Cas9 screens identify PLSCR1 as a potent anti-SARS-CoV-2 defence factor.
From: PLSCR1 is a cell-autonomous defence factor against SARS-CoV-2 infection
a, Huh7.5 cells were treated with different concentrations of IFNs (recombinant human IFNγ (rHuIFNγ) or recombinant human IFNα2a (rHuIFNα2a)) and then infected with SARS-CoV-2 (isolate USA-WA1/2020) at a multiplicity of infection (MOI) of 1. Virus production (plaque-forming units (PFU) per ml) was quantified by plaque assay at 2 days post-infection (dpi) (n = 3). b, Representative images showing infection with SARS-CoV-2 expressing mNeonGreen (SARS-CoV-2-mNG, isolate USA-WA1/2020) in negative control (NC) or STAT1-KO Huh7.5 cells in resting or IFNγ (8 U ml−1)-activated conditions. Green, SARS-CoV-2-mNG; blue, Hoechst. c, Schema showing the genome-wide CRISPR screening workflow. d, FACS plots of resting or IFNγ-activated Huh7.5 (8 U ml−1) (left) and A549-ACE2 (70 U ml−1) (right) cells infected with SARS-CoV-2-mNG at an MOI of 1 for 48 h or an MOI of 0.3 for 24 h, respectively. The percentage of infected cells is shown for populations with a high (mNGhigh) or low (mNGlow) level of mNG expression. e, Comparisons of gene-level enrichment scores (mNGhigh versus mNGlow populations) between untreated and IFNγ-treated conditions in Huh7.5 (left) and A549-ACE2 (right) cells. f, SARS-CoV-2-mNG fluorescent intensity (normalized to cell counts) in Huh7.5 (left) and average restriction ratio (−IFNγ/+IFNγ) in Huh7.5 cells of the indicated genotypes (right) (n = 3). g, SARS-CoV-2-mNG fluorescent intensity in A549-ACE2 cells of the indicated genotypes. The fluorescent intensity of mNeonGreen was quantified at 2 dpi for Huh7.5 cells (f) and 1 dpi for A549-ACE2 cells (g). Three PLSCR1-KO cell lines were generated using different sgRNAs (n = 3). Data are mean ± s.d. P values from one-way ANOVA followed by Tukey’s multiple comparison test in f,g. Scale bars (b), 500 μm. Experiments performed three times.
