V-SWITCH: A single-vector OFF-to-ON fluorescent reporter of live RNA virus infections

V-SWITCH enables OFF-to-ON mNeonGreen fluorescence in living cells during infection, with viral protease cleavage driving nuclear reconstitution across Dengue virus, Zika virus, West Nile virus, and Human Coronavirus OC43.

• Why it matters: Viral fluorescent reporters are needed to track infection dynamics and host-pathogen interactions in real time, but modular, single-vector systems that work across multiple viruses remain limited.
• What they did: The authors built a single-vector split mNeonGreen release-and-capture reporter: mNG3A(1-10) anchored to the endoplasmic reticulum via Sec61 and mNG(11) fused to BFP in the nucleus, with virus-specific protease cleavage sites swapped by restriction sites.
• The result: Infection activates fluorescence, allowing flow cytometry discrimination of infected versus uninfected cells and live-cell imaging of time-dependent, heterogeneous dynamics in A549, BJ-5 fibroblasts, HEK293T, and HeLa, supporting host-factor and compound screening.

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