Assembly and Entry of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2): Evaluation Using Virus-Like Particles
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Culture Conditions
2.2. Reagents and Antibodies
2.3. Generation of Expression Plasmids
2.4. VLP Production and Purification
2.5. Western Immunoblot Analysis
2.6. Binding and Entry Assay of SARS-CoV2 VLPs
2.7. Statistical Analysis
3. Results
3.1. Determinants of SARS-CoV2 VLP Assembly
3.2. Nucleocapsid Drives Efficient SARS-CoV2 VLP Production
3.3. Nucleocapsid Carboxyl-Terminal Domains (CTDs) Drive Efficient VLP Production
3.4. A SARS-CoV2 Nucleocapsid Fragment Interferes with VLP Production
3.5. HiBiT-Tagged SARS-CoV2 VLPs Assess Virus Binding and Entry Events
4. Discussion and Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Kumar, B.; Hawkins, G.M.; Kicmal, T.; Qing, E.; Timm, E.; Gallagher, T. Assembly and Entry of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2): Evaluation Using Virus-Like Particles. Cells 2021, 10, 853. https://doi.org/10.3390/cells10040853
Kumar B, Hawkins GM, Kicmal T, Qing E, Timm E, Gallagher T. Assembly and Entry of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2): Evaluation Using Virus-Like Particles. Cells. 2021; 10(4):853. https://doi.org/10.3390/cells10040853
Chicago/Turabian StyleKumar, Binod, Grant M. Hawkins, Tom Kicmal, Enya Qing, Emily Timm, and Tom Gallagher. 2021. "Assembly and Entry of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2): Evaluation Using Virus-Like Particles" Cells 10, no. 4: 853. https://doi.org/10.3390/cells10040853