This pseudotyped virus uses recombinant Vesicular Stomatitis Virus (rVSV) to carry the S protein mutated from D (Asp) to G (Gly) at site of 614 of SARS-CoV-2 (GenBank: MN908947), with 18-aa cytoplasmic tail truncation for optimal infection.

D614G mutant was found to be prevalent from April 2020 and enhances viral transmission (Cell 182, 1-16; 2020). Infection of cells with this pseudotyped virus carrying luciferase reporter results in high level luciferase activity. See titration result showing the undiluted pseudovirus generates signal ~100000-fold higher than uninfected control (cell alone as background), and ~10-fold higher than WT pseudovirus. The neutralizing activity of one control mAb was tested by using this pseudotyped virus.

As pseudovirus infectivity of rVSV without its original G is restricted to a single round of replication, the pseudotypes can be handled using BSL-2 containment practices. Applications include Anti-SARS-CoV-2 neutralizing antibody screening at high throughput, Anti-SARS-CoV-2 drug screening at high throughput and SARS-CoV-2 pseudovirus transduction of target cells for viral entry and functional studies.