Live Cell Imaging of Cell Cycle and Division

Premo™ FUCCI Cell Cycle Sensor 

Ready, Steady, Go with the Premo™ FUCCI Cell Cycle Sensor

Developed by Miyawaki and colleagues1, the fluorescence ubiquitination cell cycle indicator (FUCCI), 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.

  • Population-level, real-time cell cycle phase determination
  • Easy and efficient labeling through BacMam gene delivery system

Order Premo™ FUCCI

Above image—U2OS cells were transduced with Premo™ FUCCI Cell Cycle Sensor and allowed to divided. Images were collected over a 15-hour timeframe.
Monitor changes in cell proliferation with direct visual readout of cell cycle phase 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 color component to optimize color balance

How Premo™ FUCCI Cell Cycle Sensor Works

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 1 and 2).		 
Figure 1—Premo™ FUCCI Cell Cycle Sensor

Figure 1—Two-color cell cycle mapping using Premo™ FUCCI Cell Cycle Sensor.

 

Premo™ FUCCI Cell Cycle

Figure 2—Imaging cell cycle progression in live cells with Premo™ FUCCI Cell Cycle Sensor.  U2OS cells were transduced with Premo™ FUCCI Cell Cycle Sensor and allowed to divided. Images were collected over a 15-hour timeframe.
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1.  Sakaue-Sawano et al. (2008) Cell 132:487–498.