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Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals

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

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
2019
VL / 141 - BP / 4526 - EP / 4530
abstract
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.

AccesS level

Green accepted

MENTIONS DATA