A chemigenetic fluorescence lifetime-based biosensor for imaging of ERK activity
- 1ETH Zurich
Abstract
Extracellular signal-regulated kinase (ERK) governs many cellular processes, and its spatiotemporal activity dynamics are important for downstream signaling specificity. Existing fluorescent ERK activity biosensors, however, fall short in enabling spectrally multiplexed, spatially resolved, or deep tissue measurements. Here, we introduce a chemigenetic, single-fluorophore biosensor, WHaloEKAR, that reports ERK activities through changes in fluorescence lifetime. WHaloEKAR combines large lifetime changes (-0.81 ns) with the spectral versatility of the self-labeling protein HaloTag giving access to orange, red, and far-red ERK biosensors. The latter enabled the investigation of heterogeneous ERK activity at distinct subcellular locations in cultured cell lines and primary cortical rat neurons via fluorescence lifetime imaging microscopy (FLIM). Moreover, multiplexed FLIM measurements allowed to study ERK activity in parallel to the second messengers calcium and cyclic adenosine monophosphate. Finally, WHaloEKAR enabled the investigation of ERK activity in 3D samples as well as in live animals.
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BibTeX
@misc{dressler2026chemigenetic,
title = {{A chemigenetic fluorescence lifetime-based biosensor for imaging of ERK activity}},
author = {E. Dressler and R. S. Eapen and D. Pellanda and Y. Zhang and J. Julmi and M. A. Bhat and T. Patriarchi and D. Gilmour and M. S. Frei},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.09.757842},
url = {https://www.biorxiv.org/content/10.64898/2026.10.09.757842v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - A chemigenetic fluorescence lifetime-based biosensor for imaging of ERK activity
AU - E. Dressler
AU - R. S. Eapen
AU - D. Pellanda
AU - Y. Zhang
AU - J. Julmi
AU - M. A. Bhat
AU - T. Patriarchi
AU - D. Gilmour
AU - M. S. Frei
PY - 2026
DA - 2026/10/09
PB - bioRxiv
DO - 10.64898/2026.10.09.757842
UR - https://www.biorxiv.org/content/10.64898/2026.10.09.757842v1
AB - Extracellular signal-regulated kinase (ERK) governs many cellular processes, and its spatiotemporal activity dynamics are important for downstream signaling specificity. Existing fluorescent ERK activity biosensors, however, fall short in enabling spectrally multiplexed, spatially resolved, or deep tissue measurements. Here, we introduce a chemigenetic, single-fluorophore biosensor, WHaloEKAR, that reports ERK activities through changes in fluorescence lifetime. WHaloEKAR combines large lifetime changes (-0.81 ns) with the spectral versatility of the self-labeling protein HaloTag giving access to orange, red, and far-red ERK biosensors. The latter enabled the investigation of heterogeneous ERK activity at distinct subcellular locations in cultured cell lines and primary cortical rat neurons via fluorescence lifetime imaging microscopy (FLIM). Moreover, multiplexed FLIM measurements allowed to study ERK activity in parallel to the second messengers calcium and cyclic adenosine monophosphate. Finally, WHaloEKAR enabled the investigation of ERK activity in 3D samples as well as in live animals.
ER -