Dual-Mode FRET and BRET Sensors for Detecting cAMP
Dynamics
Posted on 2019-09-12 - 08:29
Genetically encoded fluorescent and
luminescent indicators have
revolutionized our ability to monitor physiology in real time, but
the separate development of new sensors for each of these imaging
modalities involves substantial effort and resources. Methods to rapidly
engineer multimodal sensors would, therefore, significantly accelerate
the diversification of sensors for simultaneous use in different systems
and applications. We hypothesized that the enhanced Nano-lanterns
could be incorporated into modular ratiometric sensors as an efficient
approach to creating dual-mode fluorescent-luminescent sensors. As
a proof-of-concept, we engineered an Epac1-based sensor that responds
to cyclic adenosine monophosphate binding with a greater than 80%
change in both Förster Resonance Energy Transfer and bioluminescent
resonance energy transfer (BRET) modes. We also demonstrate that our
new sensor reports cellular changes in G-protein-coupled signaling,
and that the ratiometric BRET mode is bright enough for subcutaneous
measurements in mice.
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French, Alexander
R.; Tesmer, Alexander L.; Tantama, Mathew (2019). Dual-Mode FRET and BRET Sensors for Detecting cAMP
Dynamics. ACS Publications. Collection. https://doi.org/10.1021/acsomega.9b01770