Murine Leukemia Virus (MLV)-based Coronavirus Spike-pseudotyped Particle Production and Infection

By J. Millet, Gary R. Whittaker. 2016

97 citations recorded by the source corpus.

Viral pseudotyped particles (pp) are enveloped virus particles, typically derived from retroviruses or rhabdoviruses, that harbor heterologous envelope glycoproteins on their surface and a genome lacking essential genes. These synthetic viral particles are safer surrogates of native viruses and acquire the tropism and host entry pathway characteristics governed by the heterologous envelope glycoprotein used. They have proven to be very useful tools used in research with many applications, such as enabling the study of entry pathways of enveloped viruses and to generate effective gene-delivery vectors. The basis for their generation lies in the capacity of some viruses, such as murine leukemia virus (MLV), to incorporate envelope glycoproteins of other viruses into a pseudotyped virus particle. These can be engineered to contain reporter genes such as luciferase, enabling quantification of virus entry events upon pseudotyped particle infection with susceptible cells. Here, we detail a protocol enabling generation of MLV-based pseudotyped particles, using the Middle East respiratory syndrome coronavirus (MERS-CoV) spike (S) as an example of a heterologous envelope glycoprotein to be incorporated. We also describe how these particles are used to infect susceptible cells and to perform a quantitative infectivity readout by a luciferase assay.

Subfields: Genetics, Infectious Diseases

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Citations in the corpus

Elicitation of Potent Neutralizing Antibody Responses by Designed Protein Nanoparticle Vaccines for SARS-CoV-2 (2020, positive): “Pseudovirus production MLV-based SARS-CoV-2 S, SARS-CoV S, and WIV-1 pseudotypes were prepared as previously described (Millet and Whittaker, 2016; Walls et al., 2020).”
Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein (2020, methodology): “Pseudovirus production MLV-based SARS-CoV S, SARS-CoV-2 S, and SARS-CoV-2 Sfur/mut pseudotypes were prepared as previously described (Millet and Whittaker, 2016).”
Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion (2020, unknown): “MLV-based SARS-CoV S-pseudotyped were prepared as previously described (Millet andWhittaker, 2016).”
Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays (2020, background): “Spike from several coronaviruses can be “pseudotyped” onto safer non-replicative viral particles in place of their endogenous entry protein, thereby making entry of these particles into cells dependent on Spike [29– 36].”