概要: Identification of host genes essential for SARS-CoV-2 infection may reveal novel therapeutic targets and inform our understanding of COVID-19 pathogenesis. Here we performed a genome-wide CRISPR screen with SARS-CoV-2 in Vero-E6 cells, identifying known SARS-CoV-2 host factors including the receptor ACE2 and protease Cathepsin L. We additionally discovered novel pro-viral genes and pathways including the SWI/SNF chromatin remodeling complex, the alarmin HMGB1, and the transcription factors Smad3 and Smad4. Small molecule inhibitors of these pathways inhibited SARS-CoV-2-infection in both monkey and human cells, demonstrating the conserved role of these genetic hits across species. We also revealed that HMGB1 is a novel regulator of ACE2 expression and critical for SARS-CoV-2 replication. In contrast, loss of the histone H3.3 chaperone complex sensitized cells to virus-induced death. Together this study reveals potential therapeutic targets for SARS-CoV-2 and highlights host genes that may regulate COVID-19 pathogenesis.
项目整体设计: RNAseq, ChIPseq (H3K4me3, H3K27ac) and ATACseq of Vero-E6 cell lines in control sgRNA and HMGB1 KO sgRNA conditions.
Vero-E6 cells were cultured in Dulbecco Modified Eagle Medium (DMEM) with 10% heat-inactivated fetal bovine serum (FBS), and 1% Penicillin/Streptomycin.
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Cells were infected with SARS-CoV-2 at a MOI of 1 for 24 hours then RNAs were extracted using Direct-zol RNA MiniPrep Kit and submitted to the Yale Center for Genome Analysis for library preparation.
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Libraries were prepared using TruSeq stranded mRNA library prep kit (Illumina)
测序信息
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poly(A)+ RNA
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PAIRED
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-
测序平台:
ILLUMINA
测序仪型号:
Illumina NovaSeq 6000
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Unspecific
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文章
Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection.
