Visualization of herpes simplex virus type 1 virions using fluorescent colors

Area of Science:

• Virology
• Molecular Biology
• Biochemistry

Background:

• Herpes simplex virus type 1 (HSV-1) is a significant human pathogen.
• Tracking viral entry and replication is crucial for understanding infection.
• Previous work engineered enhanced green fluorescent protein (eGFP) fusions to HSV-1 capsid protein VP26.

Purpose of the Study:

• To expand the palette of fluorescent tags available for HSV-1 VP26.
• To enable simultaneous tracking of multiple viral particles using different colors.
• To investigate the impact of fluorescent protein fusions on viral replication and capsid association.

Main Methods:

• Generation of HSV-1 strains with VP26 fused to various fluorescent proteins (Cerulean, Venus, mOrange, tdTomato, mCherry, Dronpa3).
• Assessment of viral replication properties in cell culture.
• Localization studies to confirm stable association of fluorescent tags with intranuclear capsids and mature virions.

Main Results:

• Successfully generated HSV-1 viruses expressing VP26 fused to Cerulean, Venus, mOrange, tdTomato, mCherry, and Dronpa3.
• These VP26-fluorescent protein fusions did not impair viral replication.
• Fusion tags remained stably associated with viral capsids and virions.
• Viruses with fluorescent proteins prone to dimerization could not be isolated.

Conclusions:

• A diverse range of fluorescently tagged HSV-1 VP26 proteins can be generated.
• These novel fluorescent tags are suitable for tracking HSV-1 during infection without affecting viral fitness.
• The ability to use multiple colors simultaneously will enhance studies of HSV-1 entry and replication dynamics.