Genetically encoded dinitrobenzyl photostabilizer for enhancing the photostability of fluorescent proteins
- 1Department of Chemistry, College of Science, Guangdong Provincial Key Laboratory of Catalysis, Guangming Advanced Research Institute, Southern University of Science and Technology Shenzhen 518055 China
- 2Taizhou Research Institute, Southern University of Science and Technology Taizhou 318001 China
Abstract
Fluorescent proteins (FPs) are essential tools for biological imaging, but their application in long-term and super-resolution microscopy is often limited by photobleaching. Inspired by “self-healing” organic dyes, we report a genetically encoded intramolecular photostabilization strategy to improve the ability of fluorescent proteins to resist photobleaching. We designed a dinitrobenzene-based unnatural amino acid (UAA), 2-NBAK, to function as an efficient photostabilizer. Using an engineered orthogonal translation system, we performed a systematic spatial scan of 192 insertion sites across the mNeonGreen (mNG) β-barrel. Our screening revealed that optimal mNG-2-NBAK variants at specific outward-pointing side chains are proposed to quench the triplet state without disrupting the chromophore emission spectra. In vitro characterization and further live-cell confocal imaging demonstrated that the optimal variants exhibited more than a twofold extension of the fluorescence half-life under intense continuous irradiation. By transplanting the optimal mNG insertion sites into another FP, we established a proof-of-concept for enhancing green FP photostability, providing a genetically encoded framework adaptable to other spectral variants with energetically matched photostabilizers.
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BibTeX
@article{li2026genetically,
title = {{Genetically encoded dinitrobenzyl photostabilizer for enhancing the photostability of fluorescent proteins}},
author = {Guihua Li and Xia Wang and Jiben Xue and Jie Wang},
journal = {RSC Chemical Biology},
year = {2026},
month = sep,
doi = {10.1039/d6cb00168h},
url = {https://doi.org/10.1039/d6cb00168h},
note = {CC BY-NC 3.0},
}
RIS
TY - JOUR
TI - Genetically encoded dinitrobenzyl photostabilizer for enhancing the photostability of fluorescent proteins
AU - Guihua Li
AU - Xia Wang
AU - Jiben Xue
AU - Jie Wang
PY - 2026
DA - 2026/09/29
JO - RSC Chemical Biology
DO - 10.1039/d6cb00168h
UR - https://doi.org/10.1039/d6cb00168h
AB - Fluorescent proteins (FPs) are essential tools for biological imaging, but their application in long-term and super-resolution microscopy is often limited by photobleaching. Inspired by “self-healing” organic dyes, we report a genetically encoded intramolecular photostabilization strategy to improve the ability of fluorescent proteins to resist photobleaching. We designed a dinitrobenzene-based unnatural amino acid (UAA), 2-NBAK, to function as an efficient photostabilizer. Using an engineered orthogonal translation system, we performed a systematic spatial scan of 192 insertion sites across the mNeonGreen (mNG) β-barrel. Our screening revealed that optimal mNG-2-NBAK variants at specific outward-pointing side chains are proposed to quench the triplet state without disrupting the chromophore emission spectra. In vitro characterization and further live-cell confocal imaging demonstrated that the optimal variants exhibited more than a twofold extension of the fluorescence half-life under intense continuous irradiation. By transplanting the optimal mNG insertion sites into another FP, we established a proof-of-concept for enhancing green FP photostability, providing a genetically encoded framework adaptable to other spectral variants with energetically matched photostabilizers.
SN - 2633-0679
ER -