VSV-LACV HAP1 human surfacesome Crisp Screen sequence reads

To identify host cell factors required for LACV entry, we used a lentiviral library of 4,630 single guide RNAs targeting 1,146 human genes that encode proteins predicted to be present on the plasma membrane, and performed a CRISPR Cas9 screen in a haploid human cell line. For this screen, we took advantage of the rapid cytopathogenicity of vesicular stomatitis virus and generated a VSV LACV chimera in which the native attachment and fusion glycoprotein of VSV was replaced with the M segment open reading frame of LACV. Gene-edited HAP1 cells were inoculated with VSV LACV at a multiplicity of infection of 3, and genomic DNA from surviving cells was sequenced to identify enriched sgRNAs. The top hit from the screen was Jagged 2, a canonical Notch ligand and single-pass type I membrane protein. Two independent biological repeats were performed, and the raw data have been submitted.