Quantitative Cell Cycle Analysis Based on an Endogenous All-in-One Reporter for Cell Tracking and Classification.
Date
2017-05-30ICR Author
Author
Zerjatke, T
Gak, IA
Kirova, D
Fuhrmann, M
Daniel, K
Gonciarz, M
Müller, D
Glauche, I
Mansfeld, J
Type
Journal Article
Metadata
Show full item recordAbstract
Cell cycle kinetics are crucial to cell fate decisions. Although live imaging has provided extensive insights into this relationship at the single-cell level, the limited number of fluorescent markers that can be used in a single experiment has hindered efforts to link the dynamics of individual proteins responsible for decision making directly to cell cycle progression. Here, we present fluorescently tagged endogenous proliferating cell nuclear antigen (PCNA) as an all-in-one cell cycle reporter that allows simultaneous analysis of cell cycle progression, including the transition into quiescence, and the dynamics of individual fate determinants. We also provide an image analysis pipeline for automated segmentation, tracking, and classification of all cell cycle phases. Combining the all-in-one reporter with labeled endogenous cyclin D1 and p21 as prime examples of cell-cycle-regulated fate determinants, we show how cell cycle and quantitative protein dynamics can be simultaneously extracted to gain insights into G1 phase regulation and responses to perturbations.
Collections
Subject
Animals
Humans
Mice
DNA Damage
Cyclins
Proliferating Cell Nuclear Antigen
Cell Cycle
G1 Phase
Cell Survival
Kinetics
Genes, Reporter
Cyclin-Dependent Kinase Inhibitor p21
Cell Tracking
Research team
Post-translational modifications and cell proliferation
Language
eng
Date accepted
2017-05-04
License start date
2017-05
Citation
Cell reports, 2017, 19 (9), pp. 1953 - 1966
Publisher
CELL PRESS