TY - JOUR
T1 - Far-red fluorescent tags for protein imaging in living tissues
AU - Shcherbo, Dmitry
AU - Murphy, Christopher S.
AU - Ermakova, Galina V.
AU - Solovieva, Elena A.
AU - Chepurnykh, Tatiana V.
AU - Shcheglov, Aleksandr S.
AU - Verkhusha, Vladislav V.
AU - Pletnev, Vladimir Z.
AU - Hazelwood, Kristin L.
AU - Roche, Patrick M.
AU - Lukyanov, Sergey
AU - Zaraisky, Andrey G.
AU - Davidson, Michael W.
AU - Chudakov, Dmitry M.
PY - 2009/3/15
Y1 - 2009/3/15
N2 - 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.
AB - 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.
KW - Far-red fluorescence
KW - Fluorescent protein
KW - Fusion protein
KW - Photostability
KW - Whole-body imaging
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U2 - 10.1042/BJ20081949
DO - 10.1042/BJ20081949
M3 - Article
C2 - 19143658
AN - SCOPUS:62149150302
SN - 0264-6021
VL - 418
SP - 567
EP - 574
JO - Biochemical Journal
JF - Biochemical Journal
IS - 3
ER -