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- Title
iGLuc: a luciferase-based inflammasome and protease activity reporter.
- Authors
Bartok, Eva; Bauernfeind, Franz; Khaminets, Maria G; Jakobs, Christopher; Monks, Brian; Fitzgerald, Katherine A; Latz, Eicke; Hornung, Veit
- Abstract
Measurement of protease activity in living cells or organisms remains a challenging task. We here present a transgene-encoded biosensor that reports the proteolytic activity of caspase-1 in the course of inflammasome activation and that of other proteases in a highly sensitive and specific manner. This protease reporter is based on the biological activity of a pro-interleukin (IL)-1?-Gaussia luciferase (iGLuc) fusion construct, in which pro-IL-1?-dependent formation of protein aggregates renders GLuc enzyme inactive. Cleavage leads to monomerization of this biosensor protein, resulting in a strong gain in luciferase activity. Exchange of the canonical caspase-1 cleavage site in this reporter construct allows the generation of protease biosensors with additional specificities. The high sensitivity, signal-to-background ratio and specificity of the iGLuc system renders it a useful tool to study proteolytic events in mouse and human cells at high throughput and to monitor protease activity in mice in vivo.
- Subjects
LUCIFERASES; INTERLEUKINS; BIOSENSORS; LABORATORY mice; PROTEOLYTIC enzyme kinetics; GENE fusion; CASPASES
- Publication
Nature Methods, 2013, Vol 10, Issue 2, p147
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth.2327