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Bright monomeric red fluorescent protein with an extended fluorescence lifetime

Abstract

Fluorescent proteins have become extremely popular tools for in vivo imaging and especially for the study of localization, motility and interaction of proteins in living cells. Here we report TagRFP, a monomeric red fluorescent protein, which is characterized by high brightness, complete chromophore maturation, prolonged fluorescence lifetime and high pH-stability. These properties make TagRFP an excellent tag for protein localization studies and fluorescence resonance energy transfer (FRET) applications.

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Figure 1: TagRFP properties.
Figure 2: TagRFP in living cells.

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Acknowledgements

We are grateful to N.E. Yelina for the critical reading of the manuscript and to L. van Weeren for technical assistance. We thank R. Tsien (University of California, San Diego) for the kind gift of mCherry. This work was supported by grants from Howard Hughes Medical Institute grant HHMI 55005618, from Molecular and Cell Biology Program RAS, from EC FP-6 Integrated Project LSHG-CT-2003-503259 and from the US National Institutes of Health (GM070358). D.M.C. and K.A.L. are supported by Grants of the President of Russian Federation MK-8236.2006.4 and Russian Science Support Foundation.

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Contributions

E.M.M., D.S., M.E.B., A.F.F. and A.G. cloned eqFP578, developed TurboRFP and TagRFP, and characterized the proteins in vitro and in living cells. J.G. and T.W.J.G. performed fluorescence lifetime imaging microscopy measurements, FRET calculations and contributed to the writing of the manuscript. A.S. performed gel-filtration experiments. K.A.L., S.L. and D.M.C. designed and planned the project, and wrote the manuscript.

Corresponding author

Correspondence to Dmitriy M Chudakov.

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Competing interests

TurboRFP and TagRFP are property of Evrogen JSC (Moscow, Russia).

K.A.L., S.L. and D.M.C. have interest in Evrogen JSC.

E.M.M. and A.G are employed by Evrogen JSC.

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Supplementary Figures 1–4, Supplementary Table 1, Supplementary Methods, Supplementary Data 1–2 (PDF 680 kb)

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Merzlyak, E., Goedhart, J., Shcherbo, D. et al. Bright monomeric red fluorescent protein with an extended fluorescence lifetime. Nat Methods 4, 555–557 (2007). https://doi.org/10.1038/nmeth1062

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