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A highly sensitive genetically encoded red cAMP sensor for multiplex imaging in vivo
Liang Wang1,2, Lingling Li3, Xuelin Li4,5
1Research Center for Primate Neuromodulation and Neuroimaging, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China. wangliang@suat-sz.edu.cn.
Nature Communications
|May 25, 2026
Summary
Researchers developed R-Flamp1, a novel red cyclic adenosine monophosphate (cAMP) sensor. This high-performance sensor overcomes limitations of existing tools, enabling clearer visualization of cAMP signaling in complex biological processes.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Circularly permuted green fluorescent protein (cpGFP)-based sensors offer high spatiotemporal resolution for monitoring cyclic adenosine monophosphate (cAMP) dynamics.
- Existing red cAMP sensors have limitations including narrow dynamic range, low brightness, aggregation, and photoactivation, hindering multiplexed imaging.
Purpose of the Study:
- To develop a red cyclic adenosine monophosphate (cAMP) sensor with improved performance characteristics.
- To overcome spectral overlap issues associated with green fluorescent protein (GFP)-based sensors in multiplexed imaging applications.
Main Methods:
- Development and characterization of a novel red cAMP sensor, R-Flamp1.
- Assessment of dynamic range, cellular brightness, cAMP affinity, response kinetics, and photoactivation properties.
- Application of R-Flamp1 in two-photon imaging and fiber photometry to visualize cAMP dynamics during behavior.
Main Results:
- R-Flamp1 exhibits a large dynamic range (>10-fold), enhanced cellular brightness, suitable cAMP affinity (Kd ~1.9 μM), and subsecond response kinetics.
- The sensor shows minimal photoactivation under blue or cyan light exposure, improving compatibility with optogenetic tools.
- R-Flamp1 enabled visualization of region-specific cAMP dynamics and revealed differential activation patterns with other signaling molecules during behavior.
Conclusions:
- R-Flamp1 is a high-performance red cAMP sensor that overcomes limitations of previous sensors, offering significant advantages for multiplexed imaging.
- This sensor provides valuable insights into the role of cAMP signaling in complex behaviors by enabling simultaneous monitoring with other indicators.
