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Rational design of a monomeric and photostable far‐red fluorescent protein for fluorescence imaging in vivo
Author(s) -
Yu Dan,
Dong Zhiqiang,
Gustafson William Clay,
RuizGonzález Rubén,
Signor Luca,
Marzocca Fanny,
Borel Franck,
Klassen Matthew P.,
Makhijani Kalpana,
Royant Antoine,
Jan YuhNung,
Weiss William A.,
Guo Su,
Shu Xiaokun
Publication year - 2016
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2843
Subject(s) - green fluorescent protein , zebrafish , fluorescence , fluorescent protein , fusion protein , microbiology and biotechnology , bimolecular fluorescence complementation , centrosome , biophysics , protein subcellular localization prediction , chemistry , far red , live cell imaging , biology , biochemistry , cell , recombinant dna , gene , physics , quantum mechanics , cell cycle , red light , botany
Fluorescent proteins (FPs) are powerful tools for cell and molecular biology. Here based on structural analysis, a blue‐shifted mutant of a recently engineered monomeric infrared fluorescent protein (mIFP) has been rationally designed. This variant, named iBlueberry, bears a single mutation that shifts both excitation and emission spectra by approximately 40 nm. Furthermore, iBlueberry is four times more photostable than mIFP, rendering it more advantageous for imaging protein dynamics. By tagging iBlueberry to centrin, it has been demonstrated that the fusion protein labels the centrosome in the developing zebrafish embryo. Together with GFP‐labeled nucleus and tdTomato‐labeled plasma membrane, time‐lapse imaging to visualize the dynamics of centrosomes in radial glia neural progenitors in the intact zebrafish brain has been demonstrated. It is further shown that iBlueberry can be used together with mIFP in two‐color protein labeling in living cells and in two‐color tumor labeling in mice.