StayRose: A photostable StayGold derivative redshifted by genetic code expansion

Photobleaching of fluorescent proteins often limits the acquisition of high-quality images in microscopy. StayGold, a novel dimeric GFP recently monomerized through sequence engineering, addresses this challenge with its high photostability. There is now a focus on producing different colored StayGold derivatives to facilitate concurrent tagging of multiple targets. The unnatural amino acid 3-aminotyrosine has previously been shown to redshift superfolder GFP upon incorporation into its chromophore via genetic code expansion. Here, we apply the same strategy to redshift StayGold through substitution of tyrosine-58 with 3-aminotyrosine. The resultant red fluorescent protein, StayRose, shows an excitation wavelength maximum of 530 nm and an emission wavelength maximum of 588 nm. Importantly, the monomeric mStayRose retains the favorable photostability in vivo in Escherichia coli and zebrafish embryos. A high-resolution crystal structure of StayRose confirms the modified structure of the amino chromophore within an unperturbed 3D fold. Although reliant on genetic code expansion, StayRose provides an important step toward developing redshifted StayGold derivatives.

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Publication Details

Journal
Journal of Biological Chemistry
Published
2025-10-17
DOI
https://doi.org/10.1016/j.jbc.2025.110832
Primary Topic
Photochromic and Fluorescence Chemistry
Type
article
Field-Weighted Citation Impact
0.00

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article

StayRose: A photostable StayGold derivative redshifted by genetic code expansion

Mohan K. Balasubramanian, Matthew Parent, Pawel K Lysyganicz, Teresa Massam-Wu et al.
Journal of Biological Chemistry
Photochromic and Fluorescence Chemistry
article

StayRose: A photostable StayGold derivative redshifted by genetic code expansion

Mohan K. Balasubramanian, Matthew Parent, Pawel K Lysyganicz, Teresa Massam-Wu, Masanori Mishima, Lijiang Song, Falk Schneider, Dipayan Akhuli, Allister Crow, Jonathan Cook, William Scott, Esther Ivorra-Molla, Abhishek Kumar, Rylie Walsh
article en

Abstract

Photobleaching of fluorescent proteins often limits the acquisition of high-quality images in microscopy. StayGold, a novel dimeric GFP recently monomerized through sequence engineering, addresses this challenge with its high photostability. There is now a focus on producing different colored StayGold derivatives to facilitate concurrent tagging of multiple targets. The unnatural amino acid 3-aminotyrosine has previously been shown to redshift superfolder GFP upon incorporation into its chromophore via genetic code expansion. Here, we apply the same strategy to redshift StayGold through substitution of tyrosine-58 with 3-aminotyrosine. The resultant red fluorescent protein, StayRose, shows an excitation wavelength maximum of 530 nm and an emission wavelength maximum of 588 nm. Importantly, the monomeric mStayRose retains the favorable photostability in vivo in Escherichia coli and zebrafish embryos. A high-resolution crystal structure of StayRose confirms the modified structure of the amino chromophore within an unperturbed 3D fold. Although reliant on genetic code expansion, StayRose provides an important step toward developing redshifted StayGold derivatives.

Journal of Biological ChemistryVol. 301(12)
Marine Biological Laboratory (US), University of Warwick (GB)
Gordon and Betty Moore Foundation, Wellcome Trust, Leverhulme Trust, University of Warwick, Medical Research Council, Engineering and Physical Sciences Research Council, Biotechnology and Biological Sciences Research Council
Openalex Percentile: Top 77%
Photochromic and Fluorescence Chemistry
DOIOpen Access PDFOpenAlex
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