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Bright far-red fluorescent protein for whole-body imaging

Abstract

For deep imaging of animal tissues, the optical window favorable for light penetration is in near-infrared wavelengths, which requires proteins with emission spectra in the far-red wavelengths. Here we report a far-red fluorescent protein, named Katushka, which is seven- to tenfold brighter compared to the spectrally close HcRed or mPlum, and is characterized by fast maturation as well as a high pH-stability and photostability. These unique characteristics make Katushka the protein of choice for visualization in living tissues. We demonstrate superiority of Katushka for whole-body imaging by direct comparison with other red and far-red fluorescent proteins. We also describe a monomeric version of Katushka, named mKate, which is characterized by high brightness and photostability, and should be an excellent fluorescent label for protein tagging in the far-red part of the spectrum.

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Figure 1: Spectral characteristics.
Figure 2: Comparison in X. laevis embryos.
Figure 3: Photobleaching experiments in living HeLa cells.

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Acknowledgements

We thank N.E. Yelina (University of East Anglia, UK) for critical reading of the manuscript. Supported by grants from Howard Hughes Medical Institute 55005618, Molecular and Cell Biology, Russian Academy of Sciences, European Commission FP-6 Integrated Project LSHG-CT-2003-503259, Russian Foundation of Basic Research 07-04-12189 and the US National Institutes of Health GM070358. D.M.C. and K.A.L. are supported by Grant of the President of Russian Federation MK-8236.2006.4, and Russian Science Support Foundation. Partially supported by grants from Howard Hughes Medical Institute 55005630, MCB Russian Academy of Sciences, Russian Foundation of Basic Research 07-04-00466 and Civilian Research and Development Foundation RUB1-2826-MO-06 to A.G.Z.

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Authors

Contributions

D.S., E.M.M., T.V.C. and A.F.F. developed Katushka and mKate and characterized the proteins in vitro and in living cells. G.V.E., E.A.S. and A.G.Z. performed experiments with X. laevis and contributed to the writing of the manuscript. E.A.B., 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

Katushka and mKate 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 T.V.C. are employed by Evrogen JSC.

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Supplementary Figures 1–6, Supplementary Methods (PDF 1455 kb)

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Shcherbo, D., Merzlyak, E., Chepurnykh, T. et al. Bright far-red fluorescent protein for whole-body imaging. Nat Methods 4, 741–746 (2007). https://doi.org/10.1038/nmeth1083

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