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Related Experiment Video

Updated: Mar 11, 2026

Analysis of Cell Cycle Position in Mammalian Cells
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Published on: January 21, 2012

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A quantitative FastFUCCI assay defines cell cycle dynamics at a single-cell level.

Siang-Boon Koh1, Patrice Mascalchi2,3, Esther Rodriguez2

  • 1Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK siangboon.koh@cantab.net fran.richards@cruk.cam.ac.uk.

Journal of Cell Science
|November 27, 2016
PubMed
Summary

A new FastFUCCI system improves live-cell imaging throughput for cell cycle analysis. This method reveals that cell death from antimitotic agents primarily occurs in interphase after mitotic slippage, not during mitosis.

Keywords:
Automated microscopyDNA replication originDrug synergyFUCCILive-cell imagingTaxane

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Drug Discovery

Background:

  • The fluorescence ubiquitination-based cell cycle indicator (FUCCI) is valuable for live-cell studies.
  • Current FUCCI assays face limitations in image processing and quantification throughput.
  • High-throughput live-cell imaging is crucial for understanding cell cycle dynamics and drug effects.

Purpose of the Study:

  • To develop a high-throughput, automated FUCCI-based assay for live-cell cycle analysis.
  • To investigate cell cycle alterations and cell death mechanisms induced by antimitotic agents.
  • To explore the potential of targeting DNA replication for sensitizing cells to chemotherapy.

Main Methods:

  • Development of a lentiviral FastFUCCI system for efficient transgene introduction.
  • Integration of FastFUCCI with an automated imaging workflow for large dataset analysis.
  • High spatiotemporal resolution analysis of single-cell responses to antimitotic drugs.

Main Results:

  • FastFUCCI improved labelling efficiency and enabled high-throughput analysis.
  • Antimitotic agents caused cell cycle arrest at G2/M, followed by cell death predominantly in interphase.
  • Mitotic slippage allowed some cells to re-enter S phase, suggesting a role for DNA replication.
  • Targeting DNA replication origin activity sensitized cells to paclitaxel.

Conclusions:

  • The FastFUCCI assay provides a powerful tool for quantifying cell cycle dynamics with high resolution.
  • Understanding cell death post-mitotic slippage is critical for evaluating drug efficacy.
  • This assay holds significant potential for preclinical drug development and cancer research.