Fluorescent Clade IIb Lineage B.1 Mpox Viruses for Antiviral Screening

Area of Science:

• Virology
• Molecular Biology
• Biotechnology

Background:

• The global mpox (monkeypox) outbreak necessitates urgent antiviral research.
• Reporter viruses are crucial for efficient antiviral drug screening.
• Monitoring mpox virus replication and spread is key to developing effective countermeasures.

Purpose of the Study:

• To engineer novel mpox clade IIb reporter viruses for enhanced antiviral screening.
• To validate the utility of these reporter viruses in tracking viral dynamics.
• To facilitate the discovery of new antiviral therapies for mpox.

Main Methods:

• Engineered two recombinant mpox clade IIb viruses with fluorescent proteins (EGFP, TurboFP635).
• Inserted fluorescent protein genes into a novel viral genome location under the vaccinia early/late promoter 7.5.
• Utilized BSC-1 cells for virus replication studies and cell flow cytometry for analysis.

Main Results:

• Recombinant viruses exhibited replication rates comparable to the parental mpox strain.
• Reporter viruses enabled clear discrimination of infected cells via flow cytometry.
• Quantified viral spread in cell culture and validated reporter system with known poxvirus inhibitors (AraC, bisbenzimide).

Conclusions:

• Developed functional mpox reporter viruses for antiviral research.
• These tools offer a robust method for monitoring mpox infection and spread.
• Reporter viruses accelerate the screening of potential mpox antiviral drugs.