Fluorescent indicators for simultaneous reporting of all four cell cycle phases

The far-red fluorescent protein mMaroon1 and a reporter based on stem-loop binding protein enables the generation of Fucci4, a 4-color cell cycle reporter system that can be used to distinguish all phases of the cell cycle. Also online, a paper by Laviv et al . uses mMaroon1 as a FRET acceptor for t...

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Published inNature methods Vol. 13; no. 12; pp. 993 - 996
Main Authors Bajar, Bryce T, Lam, Amy J, Badiee, Ryan K, Oh, Young-Hee, Chu, Jun, Zhou, Xin X, Kim, Namdoo, Kim, Benjamin B, Chung, Mingyu, Yablonovitch, Arielle L, Cruz, Barney F, Kulalert, Kanokwan, Tao, Jacqueline J, Meyer, Tobias, Su, Xiao-Dong, Lin, Michael Z
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.12.2016
Nature Publishing Group
Subjects
631/1647/1888/2249
631/1647/245/2225
631/80/2373
631/80/641
Animals
Bioinformatics
Biological Microscopy
Biological Techniques
Biomedical Engineering/Biotechnology
brief-communication
Cell Culture Techniques
Cell cycle
Cell Cycle - physiology
Chromatin - metabolism
Fluorescence
G2 Phase - physiology
HEK293 Cells
HeLa Cells
Humans
Life Sciences
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
Mice
Mitosis
Models, Molecular
Molecular Imaging - methods
NIH 3T3 Cells
Phase transitions
Proteomics
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Red Fluorescent Protein
S Phase - physiology
Time-Lapse Imaging - methods
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Summary:The far-red fluorescent protein mMaroon1 and a reporter based on stem-loop binding protein enables the generation of Fucci4, a 4-color cell cycle reporter system that can be used to distinguish all phases of the cell cycle. Also online, a paper by Laviv et al . uses mMaroon1 as a FRET acceptor for the newly developed CyRFP1. A robust method for simultaneous visualization of all four cell cycle phases in living cells is highly desirable. We developed an intensiometric reporter of the transition from S to G2 phase and engineered a far-red fluorescent protein, mMaroon1, to visualize chromatin condensation in mitosis. We combined these new reporters with the previously described Fucci system to create Fucci4, a set of four orthogonal fluorescent indicators that together resolve all cell cycle phases.
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These authors contributed equally to this work.
Current address: Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Current address: Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
ISSN:1548-7091
1548-7105
DOI:10.1038/nmeth.4045