SARS-CoV-2 Pseudotyped Virus
Mechanisms of SARS-CoV-2 infection remain poorly understood but are critical for developing therapeutic strategies to prevent viral spread. As a Biosafety Level 3 (BSL-3) agent, SARS-CoV-2 requires specialized facilities for the study of the live virus. Alternatively, it is possible to generate pseudovirus, where the structural and functional components of one virus (such as HIV, murine leukemia virus, and vesicular stomatitis virus G) are packaged as replication-defective virus particles with the SARS-CoV-2 spike (S) glycoprotein on the envelope surface. Pseudotyped virus particles can then be used to infect permissive cells that express ACE2, the cellular receptor for SARS-CoV-2, in a single round of viral entry and infection. This approach allows researchers to safely study mechanisms of infection and to determine methods of neutralizing the virus or reducing its infective potential.
The following types of viral particles have been pseudotyped with the SARS-CoV-2 Spike protein and you will find examples of each in Addgene's collection:
- HIV-based lentiviral particles
- Murine leukemia-based retroviral particles
- Vesicular stomatitis virus particles
See Addgene's Viral Plasmids and Resources page for more information.
Plasmids Expressing Spike and Spike Variants
Full-length wildtype SARS-CoV-2 spike protein is inefficient for pseudotyping; several modifications may enhance infectious particle production, including codon optimization, deletion of the C-terminal 18-21 amino acids and a D614G change which is present in many circulating variants. The table below lists spike expression plasmids that are suitable for generating SARS-CoV-2 spike pseudotyped virus using appropriate packaging plasmids. Use the search bar to find a particular modification or variant.
pBOB-CAG-SARS-CoV2-Spike-HA (Addgene plasmid #141347) is available in standard format and as a Lentiviral prep.
Envelope and Packaging Plasmids
- Popular Lentiviral Envelope and Packaging Plasmids
- Popular γ-Retrovirus Envelope and Packaging Plasmids
A few examples of reporter plasmids that can be used for measuring infectivity and neutralizing activity are listed below:
- pHAGE-CMV-Luc2-IRES-ZsGreen-W - Lentiviral dual reporter vector expressing firefly luciferase and ZsGreen.
- NL4-3 mCherry Luciferase - Lentiviral dual reporter vector expressing mCherry and firefly luciferase.
- pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral vector expressing destabilized EGFP.
- Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase with neomycin resistance.
- Bloom lab - Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein
Addgene Blog Posts
- Viral Vectors 101: Pseudotyping
- Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants