Bright Bioluminescent BRET Sensor Proteins for Measuring Intracellular Caspase Activity.

ABSTRACT: FRET-based caspase activity probes have become important tools to monitor apoptotic cell signaling. However, their dependence on external illumination is incompatible with light sensitive cells and hampers applications that suffer from autofluorescence and light scattering. Here we report the development of three caspase sensor proteins based on Bioluminescence Resonance Energy Transfer (BRET) that retain the advantages of genetically encoded, ratiometric optical probes but do not require external illumination. These sensors consist of the bright and stable luciferase NanoLuc and the fluorescent protein mNeonGreen, fused together via a linker containing a recognition site for caspase-3, -8, or -9. In vitro characterization showed that each caspase sensor displayed a robust 10-fold d

SUBMITTER: den Hamer A 

PROVIDER: S-EPMC5485374 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature