1,2,4,5-Tetrazine-tethered probes for fluorogenically imaging superoxide in live cells with ultrahigh specificity

Xuefeng Jiang, Min Li, Yule Wang, Chao Wang, Yingchao Wang, Tianruo Shen, Lili Shen, Xiaogang Liu, Yi Wang () and Xin Li ()
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Xuefeng Jiang: Zhejiang University
Min Li: Zhejiang University
Yule Wang: Zhejiang University
Chao Wang: Singapore University of Technology and Design
Yingchao Wang: Zhejiang University
Tianruo Shen: Singapore University of Technology and Design
Lili Shen: Zhejiang University
Xiaogang Liu: Singapore University of Technology and Design
Yi Wang: Zhejiang University
Xin Li: Zhejiang University

Nature Communications, 2023, vol. 14, issue 1, 1-16

Abstract: Abstract Superoxide (O2·−) is the primary reactive oxygen species in mammal cells. Detecting superoxide is crucial for understanding redox signaling but remains challenging. Herein, we introduce a class of activity-based sensing probes. The probes utilize 1,2,4,5-tetrazine as a superoxide-responsive trigger, which can be modularly tethered to various fluorophores to tune probe sensitivity and emission color. These probes afford ultra-specific and ultra-fluorogenic responses towards superoxide, and enable multiplexed imaging of various cellular superoxide levels in an organelle-resolved way. Notably, the probes reveal the aberrant superoxide generation in the pathology of myocardial ischemia/reperfusion injury, and facilitate the establishment of a high-content screening pipeline for mediators of superoxide homeostasis. One such identified mediator, coprostanone, is shown to effectively ameliorating oxidative stress-induced injury in mice with myocardial ischemia/reperfusion injury. Collectively, these results showcase the potential of 1,2,4,5-tetrazine-tethered probes as versatile tools to monitor superoxide in a range of pathophysiological settings.

Date: 2023
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DOI: 10.1038/s41467-023-37121-8

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