Pooled functional screens of viral small ORFs (smORFs) in human K562, hTERT RPE-1, and A375 cells
We designed and screened pooled libraries of small open reading frames (smORFs) encoded by human-infecting viruses. Includes an initial library of ~50,000 elements encompassing annotated coding sequences and mature peptides from viral reference genomes, together with unannotated ORFs called from ATG and CTG start codons, antisense-strand ORFs, and sequence-shuffled negative controls. A focused Enriched Library re-synthesizes hits and controls from the primary screens. Libraries were expressed from lentiviral vectors in human K562, hTERT RPE-1, and A375 cells and assayed by amplicon sequencing of element barcodes. This BioProject contains the raw sequencing data for library composition and for three classes of pooled screen: proliferation screens comparing element abundance at day 10 versus day 0; chemical ER-stress screens comparing brefeldin A- or thapsigargin-treated cells to untreated cells; and FACS-based screens for cell-surface MHC class I display, sorting cells into high and low bins with and without interferon stimulation. These data identify viral smORFs that alter human cell fitness, secretory-pathway stress tolerance, and antigen presentation, and provide a resource for functional annotation of the viral microproteome.