De novo pan-rhodamine binders for fluorescence microscopy from mammalian cells to extremophiles
8 affiliations
- 1Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, USA
- 2Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA
- 3Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA, USA
- 4Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA, USA; Department of Microbiology and Molecular Genetics, University of California, Davis, Davis, CA, USA
- 5Department of Chemistry, University of California, Riverside, Riverside, CA, USA
- 6Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA
- 7Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, USA; Department of Molecular and Chemical Sciences, Tohoku University, Sendai, Japan. Electronic address: [email protected]
- 8Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA; Biohub San Francisco, San Francisco, CA, USA. Electronic address: [email protected]
Abstract
Fluorescent imaging in live cells is a cornerstone of life sciences. While natural fluorescent proteins have been engineered to enhance individual features, no existing tag combines ideal properties into a single system: high brightness, reversible binding, compact size, and stability across diverse conditions. Here, we achieve this through de novo design of rhodamine binders (Rhobin). To harness the broad repertoire of rhodamine fluorophores, we developed a generalizable design strategy for a pan-rhodamine binder compatible with diverse wavelengths and applications. Rhobin enables live- and fixed-cell imaging of various subcellular targets in mammalian cells, showing brightness surpassing existing tags. Its reversible fluorophore binding supports super-resolution stimulated emission depletion (STED) and live-cell single-molecule imaging for extended durations compared with HaloTag. Beyond conventional systems, Rhobin enables live imaging of the extremophile Sulfolobus acidocaldarius at 75°C, previously inaccessible with current tags. Together, these results establish Rhobin as a versatile platform for next-generation imaging and biosensor design.
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BibTeX
@article{chen2026novo,
title = {{De novo pan-rhodamine binders for fluorescence microscopy from mammalian cells to extremophiles}},
author = {Yuda Chen and Klaus Yserentant and Kibeom Hong and Kehan Chen and Yiming Kuang and Robert S. Picardo and Arghya Bhowmick and Arthur Charles‐Orszag and Samuel J. Lord and Lei Lu and Kaipeng Hou and Samuel I. Mann and Sagar Bhattacharya and Maggie Horst and Jonathan B. Grimm and Luke D. Lavis and R. Dyche Mullins and William F. DeGrado and Bo Huang},
journal = {Cell},
year = {2026},
month = sep,
doi = {10.1016/j.cell.2026.08.007},
url = {https://doi.org/10.1016/j.cell.2026.08.007},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - De novo pan-rhodamine binders for fluorescence microscopy from mammalian cells to extremophiles
AU - Yuda Chen
AU - Klaus Yserentant
AU - Kibeom Hong
AU - Kehan Chen
AU - Yiming Kuang
AU - Robert S. Picardo
AU - Arghya Bhowmick
AU - Arthur Charles‐Orszag
AU - Samuel J. Lord
AU - Lei Lu
AU - Kaipeng Hou
AU - Samuel I. Mann
AU - Sagar Bhattacharya
AU - Maggie Horst
AU - Jonathan B. Grimm
AU - Luke D. Lavis
AU - R. Dyche Mullins
AU - William F. DeGrado
AU - Bo Huang
PY - 2026
DA - 2026/09/01
JO - Cell
DO - 10.1016/j.cell.2026.08.007
UR - https://doi.org/10.1016/j.cell.2026.08.007
AB - Fluorescent imaging in live cells is a cornerstone of life sciences. While natural fluorescent proteins have been engineered to enhance individual features, no existing tag combines ideal properties into a single system: high brightness, reversible binding, compact size, and stability across diverse conditions. Here, we achieve this through de novo design of rhodamine binders (Rhobin). To harness the broad repertoire of rhodamine fluorophores, we developed a generalizable design strategy for a pan-rhodamine binder compatible with diverse wavelengths and applications. Rhobin enables live- and fixed-cell imaging of various subcellular targets in mammalian cells, showing brightness surpassing existing tags. Its reversible fluorophore binding supports super-resolution stimulated emission depletion (STED) and live-cell single-molecule imaging for extended durations compared with HaloTag. Beyond conventional systems, Rhobin enables live imaging of the extremophile Sulfolobus acidocaldarius at 75°C, previously inaccessible with current tags. Together, these results establish Rhobin as a versatile platform for next-generation imaging and biosensor design.
SN - 0092-8674
SN - 1097-4172
ER -