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A FRET biosensor, SMART, monitors necroptosis in renal tubular epithelial cells in a cisplatin-induced kidney injury model

View ORCID ProfileShin Murai, Kanako Takakura, View ORCID ProfileKenta Sumimoto, View ORCID ProfileKenta Moriwaki, View ORCID ProfileKenta Terai, View ORCID ProfileSachiko Komazawa-Sakon, View ORCID ProfileYoshifumi Yamaguchi, View ORCID ProfileTetuo Mikami, View ORCID ProfileKimi Araki, Masaki Ohmuraya, View ORCID ProfileMichiyuki Matsuda, View ORCID ProfileHiroyasu Nakano
doi: https://doi.org/10.1101/2022.06.18.496655
Shin Murai
1Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori- Nishi, Ota-ku, Tokyo 143-8540, Japan
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  • ORCID record for Shin Murai
Kanako Takakura
2Imaging Platform for Spatio-Temporal Regulation, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan
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Kenta Sumimoto
3Laboratory for Mouse Genetic Engineering, RIKEN Center for Biosystems Dynamics Research, Suita 565-0874, Japan
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Kenta Moriwaki
1Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori- Nishi, Ota-ku, Tokyo 143-8540, Japan
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Kenta Terai
4Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
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Sachiko Komazawa-Sakon
1Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori- Nishi, Ota-ku, Tokyo 143-8540, Japan
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Yoshifumi Yamaguchi
5Hibernation metabolism, physiology, and development Group, Environmental Biology Division, Institute of Low Temperature Science, Hokkaido University, Kita 19, Nishi 8, Kita-ku, Sapporo, Hokkaido 060-0819, Japan
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  • ORCID record for Yoshifumi Yamaguchi
Tetuo Mikami
6Department of Pathology, Toho University School of Medicine, 5-21-16 Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan
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Kimi Araki
7Center for Animal Resources and Development, Kumamoto University, 2–2–1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan
8Center for Metabolic Regulation of Healthy Aging, Kumamoto University, 1-1-1, Honjo, Chuo-ku, Kumamoto 860-8556, Japan
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Masaki Ohmuraya
9Department of Genetics, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan
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Michiyuki Matsuda
4Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
10Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan
11Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan
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Hiroyasu Nakano
1Department of Biochemistry, Toho University School of Medicine, 5-21-16 Omori- Nishi, Ota-ku, Tokyo 143-8540, Japan
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  • ORCID record for Hiroyasu Nakano
  • For correspondence: hiroyasu.nakano@med.toho-u.ac.jp
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Abstract

Necroptosis is a regulated form of cell death involved in various pathological conditions, including ischemic reperfusion injuries, virus infections, and drug-induced tissue injuries. However, it is not fully understood when and where necroptosis occurs in vivo. We previously generated a Forster resonance energy transfer (FRET) biosensor, termed SMART (the sensor for MLKL activation based on FRET), which specifically monitored necroptosis in human and murine cell lines in vitro. Here, we generated transgenic (Tg) mice that expressed the SMART biosensor in various tissues. SMART monitored necroptosis, but not apoptosis or pyroptosis, in primary cells, including peritoneal macrophages and embryonic fibroblasts. Moreover, the FRET signal was elevated in renal tubular cells of cisplatin-treated SMART Tg mice compared to untreated SMART Tg mice. Together, SMART Tg mice may provide a valuable tool for monitoring necroptosis in different types of cells in vitro and in vivo.

Competing Interest Statement

The authors have declared no competing interest.

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Posted June 19, 2022.
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A FRET biosensor, SMART, monitors necroptosis in renal tubular epithelial cells in a cisplatin-induced kidney injury model
Shin Murai, Kanako Takakura, Kenta Sumimoto, Kenta Moriwaki, Kenta Terai, Sachiko Komazawa-Sakon, Yoshifumi Yamaguchi, Tetuo Mikami, Kimi Araki, Masaki Ohmuraya, Michiyuki Matsuda, Hiroyasu Nakano
bioRxiv 2022.06.18.496655; doi: https://doi.org/10.1101/2022.06.18.496655
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A FRET biosensor, SMART, monitors necroptosis in renal tubular epithelial cells in a cisplatin-induced kidney injury model
Shin Murai, Kanako Takakura, Kenta Sumimoto, Kenta Moriwaki, Kenta Terai, Sachiko Komazawa-Sakon, Yoshifumi Yamaguchi, Tetuo Mikami, Kimi Araki, Masaki Ohmuraya, Michiyuki Matsuda, Hiroyasu Nakano
bioRxiv 2022.06.18.496655; doi: https://doi.org/10.1101/2022.06.18.496655

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