Monitoring oxidative inflammatory processes in live cells and tissue with Hypocrates, a genetically encoded biosensor for hypochlorite

Kostyuk, A.; Tossounian, M.-A.; Panova, A.; Thauvin, M.; Wahni, K.; Van Molle, I.; Raevskii, R.; Baranov, M.; Vriz, S.; Messens, J.; Bilan, D.; Belousov, V.

doi:10.1101/2021.02.22.432222

bioRxiv · 10.1101/2021.02.22.432222

Monitoring oxidative inflammatory processes in live cells and tissue with Hypocrates, a genetically encoded biosensor for hypochlorite

Kostyuk, A.·Tossounian, M.-A.·Panova, A.·Thauvin, M.·Wahni, K.·Van Molle, I.·Raevskii, R.·Baranov, M.·Vriz, S.·Messens, J.·Bilan, D.·Belousov, V.

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Abstract

Hypochlorous acid, an aggressive oxidant, is important in immune defense against pathogens. The current lack of tools to monitor the dynamics of hypochlorous acid in live cells and tissue hinders a better understanding of inflammatory processes. We engineered a genetically encoded biosensor, Hypocrates, for the visualization of hypochlorous acid. Hypocrates consists of a circularly permuted yellow fluorescent protein integrated into the structure of the transcription repressor NemR from E. coli. We determined sensitivity, selectivity, reaction rates, and the X-ray structure of this ratiometric redox biosensor, and tested the response of Hypocrates in HeLa Kyoto cells at varying hypochlorite concentrations. By combining Hypocrates with the biosensor HyperRed, we visualized the dynamics of hypochlorous acid and hydrogen peroxide in a zebrafish tail fin injury model.

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Kostyuk, A., Tossounian, M.-A., Panova, A., Thauvin, M., Wahni, K., Van Molle, I., Raevskii, R., Baranov, M., Vriz, S., Messens, J., Bilan, D., Belousov, V.. 2021-02-22. Monitoring oxidative inflammatory processes in live cells and tissue with Hypocrates, a genetically encoded biosensor for hypochlorite. https://doi.org/10.1101/2021.02.22.432222

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