The establishment of SARS CoV-2 spike-pseudotyped lentiviral (LV) systems has enabled the rapid identification of entry inhibitors and neutralizing agents, alongside allowing for the study of this emerging pathogen in BSL-2 level facilities. While such frameworks recapitulate the cellular entry process in ACE2+ cells, they are largely unable to factor in supplemental contributions by other SARS CoV-2 genes. To address this, we performed an unbiased ORF screen and identified the nucleoprotein (N) as a potent enhancer of spike-pseudotyped LV particle infectivity. We further demonstrate that the spike protein is better enriched in virions when the particles are produced in the presence of N protein. This enrichment of spike renders LV particles more infectious as well as less vulnerable to the neutralizing effects of a human IgG-Fc fused ACE2 microbody. Importantly, this improvement in infectivity is observed with both wild-type spike protein as well as the D614G mutant. Our results hold important implications for the design and interpretation of similar LV pseudotyping-based studies.
【저자키워드】 SARS-CoV-2, nucleocapsid, Virus neutralization, spike lentiviral pseudotyping, ACE2-Fc, 【초록키워드】 SARS CoV-2, ACE2, Spike protein, Particle, pathogen, cells, Interpretation, N protein, D614G, Neutralizing, cellular entry, mutant, nucleoprotein, entry inhibitors, Entry inhibitor, Particles, SARS CoV, wild-type, virion, virions, enhancer, while, neutralizing effect, Genes, implication, BSL-2, produced, performed, ORF, less, the spike protein, fused, lentiviral, particle infectivity, 【제목키워드】 SARS CoV-2, spike, Particle, Infectivity,