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

View ORCID ProfileAlexander I. Kostyuk, View ORCID ProfileMaria-Armineh Tossounian, Anastasiya S. Panova, View ORCID ProfileMarion Thauvin, Khadija Wahni, View ORCID ProfileInge Van Molle, Roman I. Raevskii, Mikhail S. Baranov, View ORCID ProfileSophie Vriz, View ORCID ProfileJoris Messens, View ORCID ProfileDmitry S. Bilan, Vsevolod V. Belousov
doi: https://doi.org/10.1101/2021.02.22.432222
Alexander I. Kostyuk
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
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  • ORCID record for Alexander I. Kostyuk
Maria-Armineh Tossounian
3VIB-VUB Center for Structural Biology, B-1050 Brussels, Belgium
4Brussels Center for Redox Biology, B-1050 Brussels, Belgium
5Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussels, Belgium
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Anastasiya S. Panova
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
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Marion Thauvin
6Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, PSL Research University, Paris 75231, France
7Sorbonne Université, Collège Doctoral, Paris 75005, France
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Khadija Wahni
3VIB-VUB Center for Structural Biology, B-1050 Brussels, Belgium
4Brussels Center for Redox Biology, B-1050 Brussels, Belgium
5Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussels, Belgium
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Inge Van Molle
3VIB-VUB Center for Structural Biology, B-1050 Brussels, Belgium
5Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussels, Belgium
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  • ORCID record for Inge Van Molle
Roman I. Raevskii
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
8Faculty of Biology, Lomonosov Moscow State University, 119992 Moscow, Russia
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Mikhail S. Baranov
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
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Sophie Vriz
6Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, PSL Research University, Paris 75231, France
9Université de Paris, Paris 75006, France
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Joris Messens
3VIB-VUB Center for Structural Biology, B-1050 Brussels, Belgium
4Brussels Center for Redox Biology, B-1050 Brussels, Belgium
5Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussels, Belgium
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  • For correspondence: d.s.bilan@gmail.com joris.messens@vub.be belousov@fccps.ru
Dmitry S. Bilan
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
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  • For correspondence: d.s.bilan@gmail.com joris.messens@vub.be belousov@fccps.ru
Vsevolod V. Belousov
1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia
2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, 117997 Moscow, Russia
10Federal Center of Brain Research and Neurotechnologies, Federal Medical Biological Agency, 117997 Moscow, Russia
11Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073, Germany
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  • For correspondence: d.s.bilan@gmail.com joris.messens@vub.be belousov@fccps.ru
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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.

Competing Interest Statement

The authors have declared no competing interest.

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Posted February 22, 2021.
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Monitoring oxidative inflammatory processes in live cells and tissue with Hypocrates, a genetically encoded biosensor for hypochlorite
Alexander I. Kostyuk, Maria-Armineh Tossounian, Anastasiya S. Panova, Marion Thauvin, Khadija Wahni, Inge Van Molle, Roman I. Raevskii, Mikhail S. Baranov, Sophie Vriz, Joris Messens, Dmitry S. Bilan, Vsevolod V. Belousov
bioRxiv 2021.02.22.432222; doi: https://doi.org/10.1101/2021.02.22.432222
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Monitoring oxidative inflammatory processes in live cells and tissue with Hypocrates, a genetically encoded biosensor for hypochlorite
Alexander I. Kostyuk, Maria-Armineh Tossounian, Anastasiya S. Panova, Marion Thauvin, Khadija Wahni, Inge Van Molle, Roman I. Raevskii, Mikhail S. Baranov, Sophie Vriz, Joris Messens, Dmitry S. Bilan, Vsevolod V. Belousov
bioRxiv 2021.02.22.432222; doi: https://doi.org/10.1101/2021.02.22.432222

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