@article{6a00c09c0f6242b58c10da9342652283,
title = "Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals",
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.",
author = "Qiang Zhang and Antonino Schepis and Hai Huang and Junjiao Yang and Wen Ma and Joaquim Torra and Zhang, {Shao Qing} and Lina Yang and Haifan Wu and Santi Nonell and Zhiqiang Dong and Kornberg, {Thomas B.} and Coughlin, {Shaun R.} and Xiaokun Shu",
note = "Funding Information: *xiaokun.shu@ucsf.edu ORCID Haifan Wu: 0000-0002-2050-9950 Santi Nonell: 0000-0002-8900-5291 Xiaokun Shu: 0000-0001-9248-7095 Author Contributions ‡These authors contributed equally. Funding This work was supported by National Institute of General Medical Sciences (NIGMS) R01 GM115399 (to X.S.), R35 GM122548 (to T.B.K.), R01 HL054737 and R01 HL121387 (to S.R.C.), 5T32HL007731 (to H.H.), National Natural Science Foundation of China (Grant No. 31571107, 31871481 to Z.D.), and R35 GM122603 (to William F. DeGrado for support of Haifan Wu). Notes The authors declare no competing financial interest. Publisher Copyright: {\textcopyright} 2019 American Chemical Society.",
year = "2019",
month = mar,
day = "20",
doi = "10.1021/jacs.8b13042",
language = "English",
volume = "141",
pages = "4526--4530",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "11",
}