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Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting
1Department of Microbiology and Immunology, College of Veterinary Medicine,Cornell University, 2INRA, Virologie et Immunologie Moléculaires, 3Robert Frederick Smith School of Chemical and Biomolecular Engineering,Cornell University, 4Department of Microbiology, College of Agricultural and Life Sciences,Cornell University, 5Horae Gene Therapy Center,University of Massachusetts Medical School
Chapters
- 00:04Title
- 00:41Cell Seeding for Pseudotyped Particle Production
- 02:05Three-plasmid Co-transfection
- 03:53Pseudotyped Particles Collection
- 04:46Pseudotyped Particle Infection
- 05:54Infectivity Quantification by Luciferase Assay Readout
- 07:04Results: Infectivity Assays of SARS-CoV S and MERS-CoV S Pseudotyped Particles
- 08:02Conclusion
Here, we present a protocol to generate pseudotyped particles in a BSL-2 setting incorporating the spike protein of the highly pathogenic viruses Middle East respiratory syndrome and severe acute respiratory syndrome coronaviruses. These pseudotyped particles contain a luciferase reporter gene allowing quantification of virus entry into target host cells.
