PUBLICATION

Rational design of a monomeric and photostable far-red fluorescent protein for fluorescence imaging in vivo

Authors
Yu, D., Dong, Z., Gustafson, W.C., Ruiz-González, R., Signor, L., Marzocca, F., Borel, F., Klassen, M.P., Makhijani, K., Royant, A., Jan, Y.N., Weiss, W.A., Guo, S., Shu, X.
ID
ZDB-PUB-151110-10
Date
2016
Source
Protein science : a publication of the Protein Society   25(2): 308-15 (Journal)
Registered Authors
Dong, Zhiqiang, Guo, Su
Keywords
none
MeSH Terms
  • Animals
  • Cell Line, Tumor
  • Drosophila/ultrastructure
  • Female
  • Fluorescent Dyes/analysis*
  • Fluorescent Dyes/metabolism
  • HEK293 Cells
  • Humans
  • Luminescent Proteins/analysis*
  • Luminescent Proteins/genetics
  • Mice
  • Mice, Inbred BALB C
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Mutation
  • Neoplasms/diagnosis
  • Optical Imaging/methods*
  • Photobleaching
  • Protein Conformation
  • Protein Stability
  • Zebrafish/embryology
PubMed
26549191 Full text @ Protein Sci.
Abstract
Fluorescent proteins (FPs) are powerful tools for cell and molecular biology. Here we report that based on structural analysis, we have rationally designed a blue-shifted mutant of a recently engineered monomeric infrared fluorescent protein (mIFP). This variant, named iBlueberry, bears a single mutation that shifts both excitation and emission spectra by ∼40 nm. Furthermore, iBlueberry is four times more photostable than mIFP, rendering it more advantageous for imaging protein dynamics. By tagging iBlueberry to centrin, we demonstrate that the fusion protein labels the centrosome in the developing zebrafish embryo. Together with GFP-labeled nucleus and tdTomato-labeled plasma membrane, we performed time-lapse imaging to visualize the dynamics of centrosomes in radial glia neural progenitors in the intact zebrafish brain. We further show that iBlueberry can be used together with mIFP in two-color protein labeling in living cells and in two-color tumor labeling in mice. This article is protected by copyright. All rights reserved.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping