New fluorescence cell counter Biology Diagrams The cell-cycle transition from G1 to S phase has been difficult to visualize. We have harnessed antiphase oscillating proteins that mark cell-cycle transitions in order to develop genetically encoded fluorescent probes for this purpose. These probes effectively label individual G1 phase nuclei red and those in S/G2/M phases green. We were able to generate cultured cells and transgenic mice The expression levels of two closely linked proteins in the eukaryotic DNA replication network came in handy for Miyawaki's team when visualizing the cell cycle. Chromatin licensing and DNA replication factor 1 (Cdt1) "licenses" DNA to replicate during the cell cycle. Geminin inhibits Cdt1, preventing it from re-replicating DNA.
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).
Cell Cycle Indicator Biology Diagrams
By visualizing the cell cycle, Fucci is a powerful tool to investigate any process that has to do with cell growth and differentiation, such as the development and regeneration of organs as well as carcinogenesis. Cells with pure green fluorescence were either in the S or G2 phase and were distinguished by immunostaining of the S phase. The
Fucci (Fluorescent Ubiquitination-based Cell Cycle Indicator)is a set of fluorescent probes which enable the visualization of cell cycle progression in living cells. Fucci takes advantage of the fact that the replication licensing factors Cdt1 and Geminin are only present during specific phases of the cell cycle. A fusion protein of a fragment Abstract. Proper cell-cycle progression is essential for the self-renewal and differentiation of human pluripotent stem cells (hPSCs). The fluorescent ubiquitination-based cell-cycle indicator (FUCCI) has allowed the dual-color visualization of the G 1 and S/G 2 /M phases in various dynamic models, but its application in hPSCs is not widely reported. In addition, lineage-specific FUCCI
Visualizing all phases of the cell cycle Biology Diagrams
The widely used fluorescence ubiquitination cell cycle indicator (FUCCI) system, based on the expression of the cell cycle oscillators Cdt1 and geminin tagged with different fluorescent proteins
Fucci (CA) yields abundant expression of mCherry throughout G1 phase with an abrupt decrease in the red fluorescence at the end of G1 phase. Fucci (CA) can be used to reliably detect a short G1 phase and distinguish S and G2 phases, which previously was difficult to achieve. (CA) Figure1: Visualization of Cell-Cycle Progression by Fucci (SA A robust method for simultaneous visualization of all four cell cycle phases in living cells is highly desirable. We developed an intensiometric reporter of the transition from S to G2 phase and
