Simplified production of SARS-CoV-2-pseudotyped lentivirus

A common objective in developing COVID-19 therapies is to demonstrate inhibition of SARS-CoV-2 infection. Given the safety measures required in handling live SARS-CoV-2, researchers often employ alternative viruses pseudotyped with the SARS-CoV-2 spike protein (Ni et al. 2020, Shang et al. 2020). To support these efforts, we have adapted our Lenti-X packaging system to enable high-titer production of spike-pseudotyped lentiviral particles in a convenient format and developed an accompanying cell line stably expressing the human ACE2 receptor.

Packaging systems are available for the production of viral particles bearing your choice of six different SARS-CoV-2 spike protein variants:

• WT (full length)—derived from the Wuhan-Hu-1 isolate (NC_045512.2) of the novel coronavirus SARS-CoV-2, codon-optimized for expression in mammalian cells
• D614G (full length)—includes the D614G mutation in the spike protein coding sequence; the prevalence of this mutation has risen rapidly since the start of the COVID-19 pandemic (Isabel et al. 2020)
• B.1.351 (full length)—derived from SARS-CoV-2 variant originally identified in South Africa (20H/501.V2); associated with reduced susceptibility to neutralization by sera from convalescent or vaccinated individuals (Madhi et al. 2021)
• WT (truncated), D614G (truncated), and B.1.351 (truncated)—truncations of WT spike, D614G spike, and B.1.351 spike coding sequences involving removal of 19 amino acids from C-termini; the truncation is associated with increased abundance of spike protein on the resulting particles, and increased particle infectivity (Johnson et al. 2020)

For your convenience, Lenti-X SARS-CoV-2 packaging mixes are also sold separately.

References

Ni, L. et al. Detection of SARS-CoV-2-Specific Humoral and Cellular Immunity in COVID-19 Convalescent Individuals. Immunity. 52, 971–977 (2020).

Shang, J. et al. Cell entry mechanisms of SARS-CoV-2. Proc Natl Acad Sci USA. 117, 11727–11734 (2020).

Isabel, S. et al. Evolutionary and structural analyses of SARS-CoV-2 D614G spike protein mutation now documented worldwide. Sci Rep. 10, 14031 (2020).

Madhi, S. et al. Efficacy of the ChAdOx1 nCoV-19 Covid-19 Vaccine against the B.1.351 Variant. N Engl J Med. 384, 1885–1898 (2021).

Johnson, MC. et al. Optimized Pseudotyping Conditions for the SARS-COV-2 Spike Glycoprotein. J Virol. 94, e01062-20 (2020).