Details

Autor(en) / Beteiligte

• To, Tsz-Leung
• Schepis, Antonino
• Ruiz-González, Rubén
• Zhang, Qiang
• Yu, Dan
• Dong, Zhiqiang
• Coughlin, Shaun R.
• Shu, Xiaokun

Titel

Rational Design of a GFP-Based Fluorogenic Caspase Reporter for Imaging Apoptosis In Vivo

Ist Teil von

• Cell chemical biology, 2016-07, Vol.23 (7), p.875-882

Ort / Verlag

United States: Elsevier Ltd

Erscheinungsjahr

2016

Quelle

Electronic Journals Library

Beschreibungen/Notizen

• Fluorescence resonance energy transfer-based executioner caspase reporters using GFP are important tools for imaging apoptosis. While these reporters are useful for imaging apoptosis in cultured cells, their in vivo application has been handicapped by poor signal to noise. Here, we report the design and characterization of a GFP-based fluorogenic protease reporter, dubbed ZipGFP. ZipGFP-based TEV protease reporter increased fluorescence 10-fold after activation by protease. A ZipGFP-based executioner caspase reporter visualized apoptosis in live zebrafish embryos with spatiotemporal resolution. Thus, the ZipGFP-based caspase reporter may be useful for monitoring apoptosis during animal development and for designing reporters of proteases beyond the executioner caspases. •Split GFP is “zipped” or redesigned to sense protease activity•ZipGFP-based caspase reporter detects apoptosis in human cells•ZipGFP caspase reporter enables imaging physiological apoptosis in zebrafish Shu et al. develop a fluorogenic protease reporter by “zipping up” each fragment of split GFP in a manner that prevents their associate and fluorophore formation until release by specific proteolytic cleavage. The large signal enables imaging of protease activity in vivo.