SARS-CoV-2 viral budding and entry can be modeled using BSL-2 level virus-like particles

Area of Science:

• Virology
• Molecular Biology
• Biotechnology

Background:

• Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research is primarily confined to high-containment BSL-3 facilities, limiting accessibility for many laboratories.
• The global pandemic necessitates broader scientific engagement, but BSL-3 restrictions pose a significant barrier to widespread SARS-CoV-2 research.
• Developing alternative systems is crucial for facilitating research on viral mechanisms and therapeutic interventions.

Purpose of the Study:

• To create a SARS-CoV-2 virus-like particle (VLP) system for research under Biosafety Level 2 (BSL-2) conditions.
• To enable the study of SARS-CoV-2 structural protein interactions and viral life cycle steps without using infectious viral material.
• To provide a platform for evaluating potential drug inhibitors against SARS-CoV-2 entry and exit mechanisms.

Main Methods:

• Assessed the ability of the four structural proteins of SARS-CoV-2 to form virus-like particles (VLPs) in human cells.
• Developed methods for producing, purifying, and labeling these SARS-CoV-2 VLPs (fluorescently and with APEX2).
• Utilized these VLPs for studies on viral budding, entry mechanisms, and drug inhibitor screening under BSL-2 containment.

Main Results:

• Successfully produced and purified SARS-CoV-2 VLPs from human cells, demonstrating their potential for BSL-2 studies.
• Established labeling techniques (fluorescent and APEX2) for enhanced visualization and analysis of VLP interactions.
• The VLP system proved effective for evaluating viral entry processes and assessing the efficacy of drug inhibitors.

Conclusions:

• The developed SARS-CoV-2 VLP system provides a viable and accessible alternative to BSL-3 research, significantly broadening research capabilities.
• This BSL-2 compatible system facilitates the study of critical viral mechanisms like budding and entry.
• The VLP platform is a valuable resource for drug discovery and development efforts targeting SARS-CoV-2, circumventing the need for high-containment facilities.