FUCCI Reporter Gene-Based Cell Cycle Analysis

Lena Fischer; Ingo Thievessen

doi:10.1007/978-1-0716-3052-5_24

FUCCI Reporter Gene-Based Cell Cycle Analysis

Methods Mol Biol. 2023:2644:371-385. doi: 10.1007/978-1-0716-3052-5_24.

Authors

Lena Fischer¹, Ingo Thievessen²

Affiliations

¹ Biophysics Group, Department of Physics, University of Erlangen-Nuremberg, Erlangen, Germany.
² Biophysics Group, Department of Physics, University of Erlangen-Nuremberg, Erlangen, Germany. [email protected].

PMID: 37142935
DOI: 10.1007/978-1-0716-3052-5_24

Abstract

One of the most commonly assessed parameters in cellular analyses is the proliferative activity of a cell population. The fluorescence ubiquitin cell cycle indicator (FUCCI)-based system allows live and in vivo observation of cell cycle progression. Based on the mutually exclusive activity of two fluorescently labeled proteins cdt1 and geminin during the G0/1 and S/G2/M phases of the cell cycle, individual cells can be assigned to their respective cell cycle phase by fluorescence imaging of the nucleus. Here, we describe the generation of NIH/3T3 cells containing the FUCCI reporter system by lentiviral transduction and their use in 3D culture assays. The protocol can be adapted to other cell lines.

Keywords: Cdt1; Cell cycle analysis; Cell proliferation; FUCCI; Geminin; Lentivirus.

MeSH terms

Animals
Cell Cycle / genetics
Cell Cycle Proteins* / genetics
Cell Cycle Proteins* / metabolism
Cell Division
Fluorescence
Genes, Reporter
Mice
Ubiquitin*

Substances

Ubiquitin
Cell Cycle Proteins