- Zhang, Qiang;
- Schepis, Antonino;
- Huang, Hai;
- Yang, Junjiao;
- Ma, Wen;
- Torra, Joaquim;
- Zhang, Shao-Qing;
- Yang, Lina;
- Wu, Haifan;
- Nonell, Santi;
- Dong, Zhiqiang;
- Kornberg, Thomas B;
- Coughlin, Shaun R;
- Shu, Xiaokun
A family of proteases called caspases mediate apoptosis signaling in animals. We report a GFP-based fluorogenic protease reporter, dubbed "FlipGFP", by flipping a beta strand of the GFP. Upon protease activation and cleavage, the beta strand is restored, leading to reconstitution of the GFP and fluorescence. FlipGFP-based TEV protease reporter achieves 100-fold fluorescence change. A FlipGFP-based executioner caspase reporter visualized apoptosis in live zebrafish embryos with spatiotemporal resolution. FlipGFP also visualized apoptotic cells in the midgut of Drosophila. Thus, the FlipGFP-based caspase reporter will be useful for monitoring apoptosis during animal development and for designing reporters of proteases beyond caspases. The design strategy can be further applied to a red fluorescent protein for engineering a red fluorogenic protease reporter.