Protocol for characterizing the inhibition of SARS-CoV-2 infection by a protein of interest in cultured cells

Here, we present a protocol to characterize the antiviral ability of a protein of interest to SARS-CoV-2 infection in cultured cells, using MUC1 as an example. We use SARS-CoV-2 ΔN trVLP system, which utilizes transcription and replication-competent SARS-CoV-2 virus-like particles lacking nucleocapsid gene. We describe the optimized procedure to analyze protein interference of viral attachment and entry into cells, and RT-qPCR-based quantification of viral infection. The protocol can be applied to characterize more antiviral candidates and clarify their functioning stage. Graphical Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.