FUCCI Reporter Gene-Based Cell Cycle Analysis.

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.
(© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)