Bright and photostable yellow fluorescent proteins for extended imaging
Abstract Fluorescent proteins are indispensable molecular tools for visualizing biological structures and processes, but their limited photostability restricts the duration of dynamic imaging experiments. Yellow fluorescent proteins (YFPs), in particular, photobleach rapidly. Here, we introduce mGol...
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Nature Portfolio
2025-04-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-58223-5 |
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| author | Jihwan Lee Shujuan Lai Shuyuan Yang Shiqun Zhao Francisco A. Blanco Anne C. Lyons Raquel Merino-Urteaga John F. Ahrens Nathan A. Nguyen Haixin Liu Zhuohe Liu Gerard G. Lambert Nathan C. Shaner Liangyi Chen Kimberley F. Tolias Jin Zhang Taekjip Ha François St-Pierre |
| author_facet | Jihwan Lee Shujuan Lai Shuyuan Yang Shiqun Zhao Francisco A. Blanco Anne C. Lyons Raquel Merino-Urteaga John F. Ahrens Nathan A. Nguyen Haixin Liu Zhuohe Liu Gerard G. Lambert Nathan C. Shaner Liangyi Chen Kimberley F. Tolias Jin Zhang Taekjip Ha François St-Pierre |
| author_sort | Jihwan Lee |
| collection | DOAJ |
| description | Abstract Fluorescent proteins are indispensable molecular tools for visualizing biological structures and processes, but their limited photostability restricts the duration of dynamic imaging experiments. Yellow fluorescent proteins (YFPs), in particular, photobleach rapidly. Here, we introduce mGold2s and mGold2t, YFPs with up to 25-fold greater photostability than mVenus and mCitrine, two commonly used YFPs, while maintaining comparable brightness. These variants were identified using a high-throughput pooled single-cell platform, simultaneously screening for high brightness and photostability. Compared with our previous benchmark, mGold, the mGold2 variants display a ~4-fold increase in photostability without sacrificing brightness. mGold2s and mGold2t extend imaging durations across diverse modalities, including widefield, total internal reflection fluorescence (TIRF), super-resolution, single-molecule, and laser-scanning confocal microscopy. When incorporated into fluorescence resonance energy transfer (FRET)-based biosensors, the proposed YFPs enable more reliable, prolonged imaging of dynamic cellular processes. Overall, the enhanced photostability of mGold2s and mGold2t enables high-sensitivity imaging of subcellular structures and cellular activity over extended periods, broadening the scope and precision of biological imaging. |
| format | Article |
| id | doaj-art-b52af3d7c90b4ac9a5c84ac8f9d8a37e |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-b52af3d7c90b4ac9a5c84ac8f9d8a37e2025-08-20T01:54:22ZengNature PortfolioNature Communications2041-17232025-04-0116111610.1038/s41467-025-58223-5Bright and photostable yellow fluorescent proteins for extended imagingJihwan Lee0Shujuan Lai1Shuyuan Yang2Shiqun Zhao3Francisco A. Blanco4Anne C. Lyons5Raquel Merino-Urteaga6John F. Ahrens7Nathan A. Nguyen8Haixin Liu9Zhuohe Liu10Gerard G. Lambert11Nathan C. Shaner12Liangyi Chen13Kimberley F. Tolias14Jin Zhang15Taekjip Ha16François St-Pierre17Department of Neuroscience, Baylor College of MedicineDepartment of Neuroscience, Baylor College of MedicineDepartment of Chemical and Biomolecular Engineering, Rice UniversityState Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, School of Future Technology, Peking UniversityDepartment of Neuroscience, Baylor College of MedicineShu Chien-Gene Lay Department of Bioengineering, University of California San DiegoHoward Hughes Medical Institute and Program in Cellular and Molecular Medicine, Boston Children’s HospitalDepartment of Bioengineering, Rice UniversityDepartment of Biosciences, Rice UniversityDepartment of Neuroscience, Baylor College of MedicineDepartment of Neuroscience, Baylor College of MedicineDepartment of Neurosciences, University of California San Diego School of MedicineDepartment of Pharmacology, University of California San DiegoState Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, School of Future Technology, Peking UniversityDepartment of Neuroscience, Baylor College of MedicineShu Chien-Gene Lay Department of Bioengineering, University of California San DiegoHoward Hughes Medical Institute and Program in Cellular and Molecular Medicine, Boston Children’s HospitalDepartment of Neuroscience, Baylor College of MedicineAbstract Fluorescent proteins are indispensable molecular tools for visualizing biological structures and processes, but their limited photostability restricts the duration of dynamic imaging experiments. Yellow fluorescent proteins (YFPs), in particular, photobleach rapidly. Here, we introduce mGold2s and mGold2t, YFPs with up to 25-fold greater photostability than mVenus and mCitrine, two commonly used YFPs, while maintaining comparable brightness. These variants were identified using a high-throughput pooled single-cell platform, simultaneously screening for high brightness and photostability. Compared with our previous benchmark, mGold, the mGold2 variants display a ~4-fold increase in photostability without sacrificing brightness. mGold2s and mGold2t extend imaging durations across diverse modalities, including widefield, total internal reflection fluorescence (TIRF), super-resolution, single-molecule, and laser-scanning confocal microscopy. When incorporated into fluorescence resonance energy transfer (FRET)-based biosensors, the proposed YFPs enable more reliable, prolonged imaging of dynamic cellular processes. Overall, the enhanced photostability of mGold2s and mGold2t enables high-sensitivity imaging of subcellular structures and cellular activity over extended periods, broadening the scope and precision of biological imaging.https://doi.org/10.1038/s41467-025-58223-5 |
| spellingShingle | Jihwan Lee Shujuan Lai Shuyuan Yang Shiqun Zhao Francisco A. Blanco Anne C. Lyons Raquel Merino-Urteaga John F. Ahrens Nathan A. Nguyen Haixin Liu Zhuohe Liu Gerard G. Lambert Nathan C. Shaner Liangyi Chen Kimberley F. Tolias Jin Zhang Taekjip Ha François St-Pierre Bright and photostable yellow fluorescent proteins for extended imaging Nature Communications |
| title | Bright and photostable yellow fluorescent proteins for extended imaging |
| title_full | Bright and photostable yellow fluorescent proteins for extended imaging |
| title_fullStr | Bright and photostable yellow fluorescent proteins for extended imaging |
| title_full_unstemmed | Bright and photostable yellow fluorescent proteins for extended imaging |
| title_short | Bright and photostable yellow fluorescent proteins for extended imaging |
| title_sort | bright and photostable yellow fluorescent proteins for extended imaging |
| url | https://doi.org/10.1038/s41467-025-58223-5 |
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