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

Qiang Zhang, Antonino Schepis, Hai Huang, Junjiao Yang, Wen Ma, Joaquim Torra, Shao Qing Zhang, Lina Yang, Haifan Wu, Santi Nonell, Zhiqiang Dong, Thomas B. Kornberg, Shaun R. Coughlin, Xiaokun Shu

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Resum

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.

Idioma originalAnglès
Pàgines (de-a)4526-4530
Nombre de pàgines5
RevistaJournal of the American Chemical Society
Volum141
Número11
DOIs
Estat de la publicacióPublicada - 20 de març 2019

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