Premo FUCCI Cell Cycle Sensor

Premo FUCCI Cell Cycle Sensor

Developed by Miyawaki and colleagues, the Invitrogen Premo FUCCI (fluorescence ubiquitination cell cycle indicator) Cell Cycle Sensor is a genetically-encoded, two-color (red and green) indicator that lets you follow cell division within a cell population. We have incorporated the FUCCI genetic constructs into the powerful BacMam gene delivery system, creating a simple and efficient method for labeling cells and following their division.

How Premo FUCCI Cell Cycle Sensor Works

With Premo FUCCI sensor you can:

  • Monitor changes in cell proliferation with direct visual readout of cell cycle phase (Figure 1) for every cell in the population
  • Efficient BacMam transfection in wide variety of cultured, primary, and stem cell types
  • Independently control expression levels of each protein component to optimize balance

The Premo FUCCI Cell Cycle Sensor consists of a fluorescent protein-based system that employs both a red (RFP) and a green (GFP) fluorescent protein fused to different regulators of the cell cycle: cdt1 and geminin.

During the cell cycle, these two proteins are ubiquitinated by specific ubiquitin E3 ligases, which target them to the proteasome for degradation. The E3 ligases display temporal regulation of their activity which results in the biphasic cycling of geminin and cdt1levels during the cell cycle. In the G1 phase of the cell cycle, geminin is degraded; therefore, only cdt1 tagged with RFP is present and appears as red fluorescence within the nuclei. In the S, G2, and M phases, cdt1 is degraded and only geminin tagged with GFP remains, resulting in cells with green fluorescent nuclei.

During the G1/S transition, when cdt1 levels are decreasing and geminin levels increasing, both proteins are present, and when the green and red images are overlaid, nuclei appear yellow fluorescent. This dynamic color change, from red-through-yellow-to-green, represents the progression through cell cycle and division (Figures 2 and 3).

Time-lapse series of transduced U2OS cells

Figure 1. Time-lapse series of transduced U2OS cells. U2OS cells were transduced with Premo FUCCI Cell Cycle Sensor and allowed to divide. Images were collected over a 15-hour timeframe.


Table 1. Premo FUCCI Cell Cycle Sensor specifications

 
ReadoutAs a cell progresses through the cell cycle, fluorescence of the Premo FUCCI sensor changes from red to green
Common filter setRFPFITC
ReporterRFPGFP
Ex/Em (nm)555/584485/520
Live-cell compatibleYes
Stains fixed cellsNo
Formaldehyde fixableYes
MultiplexingYes
Signal-to-noise4 stars
Photostability4 stars
PlatformImaging, HCS
Bibliography 
Format2 x 200 μL2 x 1,000 μL
Cat. No.P36238P36237
BacMam technology

The BacMam system uses a modified insect cell virus (baculovirus) as a vehicle to efficiently deliver and express genes in mammalian cells with minimum effort and toxicity.

BacMam compatible cells

BacMam reagents can be used in a variety of mammalian cell types.  View the cell types that have been transduced using BacMam technology and learn how to optimize conditions for your cell type.

Resources

Tools

Flow Cytometry Panel Builder—Design your flow cytometry panel with this online tool for a simplified, customizable experience to fit your needs.

Fluorescence SpectraViewer—Online tool for visualization of the excitation and emission of fluorescent reagents. Tool allows for checking spectral compatibility for multiple fluorophores.

Cell viability, proliferation, and cell cycle information—Find educational resources such as application notes, webinars, videos, articles, and more that cover the use of many of our reagents and kits for monitoring cell function.

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