Improved SARS-CoV-2 Spike Glycoproteins for Pseudotyping Lentiviral Vectors

The spike (S) glycoprotein of SARS-Cov-2 facilitates viral entry into target cells via the cell surface receptor angiotensin-converting enzyme 2 (ACE2). Third generation HIV-1 lentiviral vectors can be pseudotyped to replace native CD4 tropic envelope protein virus and thereby either limit or expand population. We generated a modified S pseudotype which efficiently transduced ACE2-expressing with high specificity contain minimal off-target transduction ACE2 negative cells. By utilizing optimized codons, modifying cytoplasmic tail domain, including mutant form protein, we an expression plasmid encoding that produces S-pseudotyped at infectious titer (TU/mL) 1000-fold higher than unmodified 4 10-fold more specific widely used delta-19 vectors. replication-defective eliminate need for biosafety-level-3 laboratories required when developing therapeutics against SARS-CoV-2 live virus. Furthermore, activity may as tools understand development immunity SARS-CoV-2, develop assays neutralizing antibodies other agents block binding, allow in vivo imaging studies