Establishment of a Panel of Human Cell Lines to Identify Cellular Receptors Used by Enteroviruses to Infect Cells

Non-pathogenic natural and recombinant strains of human <i>Enteroviruses</i> are the subject of ongoing study with some strains having been approved for use as anticancer agents. The efficacy of oncolytic virotherapy depends upon identifying the receptor utilized by a specific strain for cell entry, and the presence of this receptor on the surface of cancer cells. Accordingly, a rapid and straightforward approach to determining the enteroviral receptors is necessary for developing an effective patient-specific, virus-based cancer therapy. To this end, we created a panel of seven lines with double knockouts on the background of the HEK293T cell line, which lacks the <i>IFNAR1</i> gene. In these lines, the main viral receptor genes, including <i>PVR</i>, <i>CXADR</i>, <i>CD55</i>, <i>ITGA2</i>, <i>SCARB2</i>, <i>ICAM1</i>, and <i>FCGRT</i>, were knocked out using the CRISPR/Cas9 system. The panel of lines was validated on twelve different <i>Enteroviruses</i> types, providing a basis for studying the molecular mechanisms of enterovirus entry into cells, and for developing new therapeutic strains.