Related Experiment Video
Updated: Mar 18, 2026
08:40
Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting
Published on: March 1, 2019
59.8K
Procedures for the Identification of SARS-CoV-2 Entry Inhibitors as Potential Antivirals using MLV-Based
Andreina Ayala1, Lissett Avecilla2, Natalia Samuel1
1Centro de Biología Celular y Molecular de enfermedades, Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT AIP).
Journal of Visualized Experiments : Jove
|March 16, 2026
Summary
A new protocol uses SARS-CoV-2 spike pseudoviruses for efficient viral entry studies. This platform aids in discovering antiviral drugs against emerging variants under safe, accessible conditions.
Area of Science:
- Virology
- Infectious Diseases
- Drug Discovery
Background:
- Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) emerged in 2019, causing significant global disease and mortality.
- Developing safe and accessible methods to study viral entry and test antiviral compounds is crucial for combating emerging variants.
Purpose of the Study:
- To describe an optimized protocol for generating SARS-CoV-2 spike pseudoviruses for viral entry studies.
- To establish a reliable platform for screening antiviral compounds against SARS-CoV-2.
Main Methods:
- Generation of murine leukemia virus (MLV)-based pseudoviruses displaying the SARS-CoV-2 spike protein and containing a luciferase reporter.
- Optimization of cell lines and high-efficiency, cost-effective transfection for pseudovirus production.
- Titration of pseudovirus infectivity and compound screening using luciferase activity and MTT viability assays.
Main Results:
- Demonstrated robust luciferase activity in cells infected with SARS-CoV-2 pseudoviruses.
- Confirmed assay specificity using control pseudoviruses (spike-less and VSV-G).
- Showcased the system's capability for compound screening, quantifying inhibition of viral entry and cytotoxicity.
Conclusions:
- The developed protocol provides a reproducible and scalable platform for medium to high-throughput screening of viral entry inhibitors.
- This system facilitates the identification of novel antiviral candidates and advances understanding of SARS-CoV-2 entry mechanisms.
- Enables viral entry studies under Biosafety Level 2 conditions, enhancing accessibility for research.
