mCranberry: A rationally engineered monomeric red fluorescent protein with 85% quantum yield
- 1The Ohio State University
Abstract
Fluorescent proteins (FPs) are the workhorse of modern-day cellular imaging and biosensing. However, rationally predicting and engineering the photophysical properties of FPs remain challenging, as these are governed by complex three-dimensional microenvironments and post-translational modifications. Here, we present a novel surface-mutation-guided engineering strategy combined with iterative rounds of rational mutagenesis to improve key photophysical properties of red fluorescent proteins (RFPs). This approach generated two bright, monomeric RFP variants from the mFruits lineage, mCranberry and mCranberry2, with quantum yields of 85% and 80%, respectively. Strategic incorporation of charged amino acid residues on the protein surface enhanced fluorescence quantum yield without compromising photostability. Comprehensive photophysical characterization and live-cell microscopy demonstrate enhanced performance of these variants and establish their utility as robust probes for fluorescence imaging. However, the exact mechanism of such improvements is subject to further investigation. Collectively, this work introduces a strategy for optimizing fluorescence properties and provides a framework that may be broadly applicable to other pigment-protein systems such as genetically encoded metal-ion indicators and photosynthetic light-harvesting complexes.
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BibTeX
@misc{chaubey2026mcranberry,
title = {{mCranberry: A rationally engineered monomeric red fluorescent protein with 85% quantum yield}},
author = {S. Chaubey and S. Mukherjee and A. Adams and A. R. Walker and M. L. M. Furgason and H. K. R. R. Babu and I. I. Mathews and P. Manna},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.01.755472},
url = {https://www.biorxiv.org/content/10.64898/2026.10.01.755472v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - mCranberry: A rationally engineered monomeric red fluorescent protein with 85% quantum yield
AU - S. Chaubey
AU - S. Mukherjee
AU - A. Adams
AU - A. R. Walker
AU - M. L. M. Furgason
AU - H. K. R. R. Babu
AU - I. I. Mathews
AU - P. Manna
PY - 2026
DA - 2026/10/01
PB - bioRxiv
DO - 10.64898/2026.10.01.755472
UR - https://www.biorxiv.org/content/10.64898/2026.10.01.755472v1
AB - Fluorescent proteins (FPs) are the workhorse of modern-day cellular imaging and biosensing. However, rationally predicting and engineering the photophysical properties of FPs remain challenging, as these are governed by complex three-dimensional microenvironments and post-translational modifications. Here, we present a novel surface-mutation-guided engineering strategy combined with iterative rounds of rational mutagenesis to improve key photophysical properties of red fluorescent proteins (RFPs). This approach generated two bright, monomeric RFP variants from the mFruits lineage, mCranberry and mCranberry2, with quantum yields of 85% and 80%, respectively. Strategic incorporation of charged amino acid residues on the protein surface enhanced fluorescence quantum yield without compromising photostability. Comprehensive photophysical characterization and live-cell microscopy demonstrate enhanced performance of these variants and establish their utility as robust probes for fluorescence imaging. However, the exact mechanism of such improvements is subject to further investigation. Collectively, this work introduces a strategy for optimizing fluorescence properties and provides a framework that may be broadly applicable to other pigment-protein systems such as genetically encoded metal-ion indicators and photosynthetic light-harvesting complexes.
ER -