Journal: Nature Genetics
Article Title: Genome-wide bidirectional CRISPR screens identify mucins as host factors modulating SARS-CoV-2 infection
doi: 10.1038/s41588-022-01131-x
Figure Lengend Snippet: a , Schematic of genome-wide CRISPR KO and activation screens for SARS-CoV-2 host factors, conducted in parallel. Calu-3 cells stably expressing Cas9 for the LOF screen or dCas9 and transcriptional activators for the GOF screen were transduced with pooled guide RNA libraries. Following infection with SARS-CoV-2, cells were harvested after at least 70% CPE was evident. Next-generation sequencing was performed to identify host factors and assign proviral and antiviral roles based on guide RNA enrichment or depletion compared with uninfected controls. b , Manhattan plot displaying the top 13 enriched genes identified in the LOF screen. c , Manhattan plot displaying the top 13 depleted genes in the GOF screen. d , Manhattan plot displaying the top 13 enriched genes in the GOF screen. All genes are ranked based on MAGeCK robust rank aggregation (RRA) score. Red dots indicate putative antiviral genes with FDR < 0.05, blue dots indicate putative proviral genes with FDR < 0.05.
Article Snippet: Approximately 24 h post-transduction, cells were selected with 10 μg ml −1 blasticidin for 10 d. Cas9 and dCas–VP64 Calu-3 knock-in lines were then transduced with Brunello and Calabrese Set A libraries (Addgene #73179 and #92379) as appropriate, which were gifts from J. Doench.
Techniques: Genome Wide, CRISPR, Activation Assay, Stable Transfection, Expressing, Transduction, Infection, Next-Generation Sequencing