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 Verkhusha, Vladimir Z. Pletnev, Kristin L. Hazelwood, Patrick M. Roche, Sergey Lukyanov, Andrey G. Zaraisky, Michael W. Davidson, Dmitry M. Chudakov

Research output: Contribution to journalArticle

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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

Fingerprint

Molecular Motor Proteins
Tissue
Imaging techniques
Whole Body Imaging
Luminance
Xenopus laevis
Fluorescent Dyes
Proteins
Embryonic Structures
Color
Fluorescence
Labeling
Toxicity
Animals
Fusion reactions
Cells
red fluorescent protein

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Shcherbo, D., Murphy, C. S., Ermakova, G. V., Solovieva, E. A., Chepurnykh, T. V., Shcheglov, A. S., ... 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

Far-red fluorescent tags for protein imaging in living tissues. / Shcherbo, Dmitry; Murphy, Christopher S.; Ermakova, Galina V.; Solovieva, Elena A.; Chepurnykh, Tatiana V.; Shcheglov, Aleksandr S.; Verkhusha, Vladislav; Pletnev, Vladimir Z.; Hazelwood, Kristin L.; Roche, Patrick M.; Lukyanov, Sergey; Zaraisky, Andrey G.; Davidson, Michael W.; Chudakov, Dmitry M.

In: Biochemical Journal, Vol. 418, No. 3, 15.03.2009, p. 567-574.

Research output: Contribution to journalArticle

Shcherbo, D, Murphy, CS, Ermakova, GV, Solovieva, EA, Chepurnykh, TV, Shcheglov, AS, Verkhusha, V, Pletnev, VZ, Hazelwood, KL, Roche, PM, Lukyanov, S, Zaraisky, AG, Davidson, MW & Chudakov, DM 2009, 'Far-red fluorescent tags for protein imaging in living tissues', Biochemical Journal, vol. 418, no. 3, pp. 567-574. https://doi.org/10.1042/BJ20081949
Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, Shcheglov AS et al. Far-red fluorescent tags for protein imaging in living tissues. Biochemical Journal. 2009 Mar 15;418(3):567-574. https://doi.org/10.1042/BJ20081949
Shcherbo, Dmitry ; Murphy, Christopher S. ; Ermakova, Galina V. ; Solovieva, Elena A. ; Chepurnykh, Tatiana V. ; Shcheglov, Aleksandr S. ; Verkhusha, Vladislav ; Pletnev, Vladimir Z. ; Hazelwood, Kristin L. ; Roche, Patrick M. ; Lukyanov, Sergey ; Zaraisky, Andrey G. ; Davidson, Michael W. ; Chudakov, Dmitry M. / Far-red fluorescent tags for protein imaging in living tissues. In: Biochemical Journal. 2009 ; Vol. 418, No. 3. pp. 567-574.
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