Far-red fluorescent tags for protein imaging in living tissues

Dmitry Shcherbo, Christopher S. Murphy, Galina V. Ermakova, Elena A. Solovieva, Tatiana V. Chepurnykh, Aleksandr S. Shcheglov, Vladislav V. Verkhusha, Vladimir Z. Pletnev, Kristin L. Hazelwood, Patrick M. Roche, Sergey Lukyanov, Andrey G. Zaraisky, Michael W. Davidson, Dmitry M. Chudakov

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316 Scopus citations

Abstract

A vast colour palette of monomeric fluorescent proteins has been developed to investigate protein localization, motility and interactions. However, low brightness has remained a problem in far-red variants, which hampers multicolour labelling and whole-body imaging techniques. In the present paper, we report mKate2, a monomeric far-red fluorescent protein that is almost 3-fold brighter than the previously reported mKate and is 10-fold brighter than mPlum. The high-brightness, far-red emission spectrum, excellent pH resistance and photostability, coupled with low toxicity demonstrated in transgenic Xenopus laevis embryos, make mKate2 a superior fluorescent tag for imaging in living tissues. We also report tdKatushka2, a tandem far-red tag that performs well in fusions, provides 4-fold brighter near-IR fluorescence compared with mRaspberry or mCherry, and is 20-fold brighter than mPlum. Together, monomeric mKate2 and pseudomonomeric tdKatushka2 represent the next generation of extrabright far-red fluorescent probes offering novel possibilities for fluorescent imaging of proteins in living cells and animals.

Original languageEnglish (US)
Pages (from-to)567-574
Number of pages8
JournalBiochemical Journal
Volume418
Issue number3
DOIs
StatePublished - Mar 15 2009

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Keywords

  • Far-red fluorescence
  • Fluorescent protein
  • Fusion protein
  • Photostability
  • Whole-body imaging

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Shcherbo, D., Murphy, C. S., Ermakova, G. V., Solovieva, E. A., Chepurnykh, T. V., Shcheglov, A. S., Verkhusha, V. V., Pletnev, V. Z., Hazelwood, K. L., Roche, P. M., Lukyanov, S., Zaraisky, A. G., Davidson, M. W., & Chudakov, D. M. (2009). Far-red fluorescent tags for protein imaging in living tissues. Biochemical Journal, 418(3), 567-574. https://doi.org/10.1042/BJ20081949