FUCCI-Red: a single-color cell cycle indicator for fluorescence lifetime imaging

Marina V. Shirmanova, Dmitry A. Gorbachev, Karen S. Sarkisyan, Alina P. Parnes, Alena I. Gavrina, Anastasia V. Polozova, Tatyana F. Kovaleva, Ludmila B. Snopova, Varvara V. Dudenkova, Elena V. Zagaynova, Konstantin A. Lukyanov

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

The phase of the cell cycle determines numerous aspects of cancer cell behaviour including invasiveness, ability to migrate and responsiveness to cytotoxic drugs. To non-invasively monitor progression of cell cycle in vivo, a family of genetically encoded fluorescent indicators, FUCCI (fluorescent ubiquitination-based cell cycle indicator), has been developed. Existing versions of FUCCI are based on fluorescent proteins of two or more different colors fused to cell-cycle-dependent degradation motifs. Thus, FUCCI-expressing cells emit light of different colors in different phases providing a robust way to monitor cell cycle progression by fluorescence microscopy and flow cytometry but limiting the possibility to simultaneously visualize other markers. To overcome this limitation, we developed a single-color variant of FUCCI, called FUCCI-Red, which utilizes two red fluorescent proteins with distinct fluorescence lifetimes, mCherry and mKate2. Similarly to FUCCI, these proteins carry cell cycle-dependent degradation motifs to resolve G1 and S/G2/M phases. We showed utility of FUCCI-Red by visualizing cell cycle progression of cancer cells in 2D and 3D cultures and monitoring development of tumors in vivo by confocal and fluorescence lifetime imaging microscopy (FLIM). Single-channel registration and red-shifted spectra make FUCCI-Red sensor a promising instrument for multiparameter in vivo imaging applications, which was demonstrated by simultaneous detection of cellular metabolic state using endogenous fluorescence in the blue range.

Original languageEnglish
Pages (from-to)3467-3476
Number of pages10
JournalCellular and Molecular Life Sciences
Volume78
Issue number7
DOIs
Publication statusPublished - Apr 2021

Keywords

  • Cell cycle
  • Fluorescence lifetime
  • Fluorescent proteins
  • Genetically encoded probes
  • Multiparameter in vivo imaging
  • Tumor model

Fingerprint

Dive into the research topics of 'FUCCI-Red: a single-color cell cycle indicator for fluorescence lifetime imaging'. Together they form a unique fingerprint.

Cite this