Anticancer drug screening in 2D/3D cultures using cell cycle marker Fucci
Fucci(CA)5 is a new variant of Fluorescent Ubiquitination-based Cell-Cycle Indicator that allows observation of cell cycle progression in live and fixed samples. The cell cycle phase is visualized by utilizing the ubiquitination domains of human Cdt1 (hCdt1(1/100)Cy(-)) fused with a red fluorescent protein (AzaleaB5), and human Geminin (hGem(1/110)) fused with a green fluorescent protein (h2-3), respectively. Because these red and green probes are ubiquitinated and degraded only in S and G1 phases, respectively, Fucci(CA)5 shows red fluorescence in G1 phase nuclei, green in S phase nuclei and yellow (red + green) in G2-M phase nuclei.
This application note presents an example using HepG2 cells that express Fucci(CA)5 to observe the response of drugs such as anticancer agents with a high content analysis system. Comparing data obtained from 2D and 3D culture conditions has emphasized the importance of understanding spatio-temporal data to study drug effects.
For further information, download the application note at the top of the page.
Reference: Sakaue-Sawano A, Miyawaki A. Mol Cell. 2017. DOI：10.1016/j.molcel.2017.10.001
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