Nsp16 shields <scp>SARS–CoV</scp> ‐2 from efficient <scp>MDA5</scp> sensing and <scp>IFIT1</scp> ‐mediated restriction

Methylation of the mRNA 5′ cap by cellular methyltransferases enables efficient translation and avoids recognition by innate immune factors. Coronaviruses encode viral 2′‐O‐methyltransferases to shield their RNA from host factors. Here, we generate recombinant SARS–CoV‐2 harboring a catalytically inactive 2′‐O‐methyltransferase Nsp16, Nsp16mut, and analyze viral replication in human lung epithelial cells. Although replication is only slightly attenuated, we find SARS–CoV‐2 Nsp16mut to be highly immunogenic, resulting in a strongly enhanced release of type I interferon upon infection. The elevated immunogenicity of Nsp16mut is absent in cells lacking the RNA sensor MDA5. In addition, we report that Nsp16mut is highly sensitive to type I IFN treatment and demonstrate that this strong antiviral effect of type I IFN is mediated by the restriction factor IFIT1. Together, we describe a dual role for the 2′‐O‐methyltransferase Nsp16 during SARS–CoV‐2 replication in avoiding efficient recognition by MDA5 and in shielding its RNA from interferon‐induced antiviral responses, thereby identifying Nsp16 as a promising target for generating attenuated and highly immunogenic SARS–CoV‐2 strains and as a potential candidate for therapeutic intervention.