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Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting
Published on: March 1, 2019
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An optimized and robust SARS-CoV-2 pseudovirus system for viral entry research
Peng Yang1, Yang Yang2, Yuming Wu2
1College of Life Sciences, Henan Normal University, Xinxiang 453007, China; Beijing Institute of Radiation Medicine, Beijing 100850, China.
Journal of Virological Methods
|June 28, 2021
Summary
Researchers developed an optimized SARS-CoV-2 pseudovirus particle (SARS2pp) system for studying Coronavirus Disease 2019 (COVID-19) entry and inhibitors in BSL-2 labs. This robust system aids in evaluating inhibitors and understanding viral mechanisms.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- SARS-CoV-2 causes COVID-19, necessitating safe laboratory models for research.
- Existing research requires efficient systems for studying viral entry and developing countermeasures.
Purpose of the Study:
- To construct and optimize a SARS-CoV-2 pseudovirus particle (SARS2pp) production and infection system for BSL-2 laboratory use.
- To enhance pseudovirus yield and establish a reliable system for evaluating SARS-CoV-2 inhibitors.
Main Methods:
- Constructed a lentiviral vector system with dual-reporter genes (eGFP and Luc2) for SARS2pp production.
- Optimized spike protein modifications, expression promoters (CMV), sequence screening, and medium additives (BSA).
- Validated the system's performance by assessing its linear relationship with multiplicity of infection (MOI) and testing inhibitors.
Main Results:
- Identified key modifications for significantly improved SARS2pp titer, including spike protein alterations and CMV promoter use.
- Demonstrated a robust linear relationship between MOI and infection across a wide range.
- Successfully utilized the system to evaluate SARS-CoV-2 inhibitors like ACE2 and neutralizing antibodies.
- Observed differential susceptibility of kidney, liver, and small intestine cell lines to SARS2pp.
Conclusions:
- The developed SARS2pp system is robust and efficient for BSL-2 research on SARS-CoV-2 entry mechanisms and inhibitors.
- This system facilitates the evaluation of antiviral drugs and antibodies.
- The platform is adaptable for studying emerging SARS-CoV-2 variants.
