Clontech’s cell cycle reporter vectors let you monitor cell cycle progression in living cells, in real-time, without fixation. They even let you visualize cell shape.
Our cell cycle reporter vectors deliver fluorescent, ubiquitination–based, cell-cycle indicators (Fucci) that allow you to identify cells in various phases of the cell cycle. These Fucci probes contain Cdt1 or Geminin, proteins whose levels fluctuate differentially throughout the cell cycle: Cdt1 levels peak in G1 phase; as cells transition into S phase, Cdt1 levels fall and Geminin levels rise, remaining high until the cells are back in G1. Cells control Cdt1 and Geminin levels post-translationally, using ubiquitination to target the unwanted proteins for proteasomal degradation. To allow Geminin and Cdt1 to be easily visualized, each is expressed with a red or cyan Living Colors fluorescent tag. These probes allow precise, visual evaluation of the cell cycle phase.
Monitor Cell Cycle and Shape in Real-Time
Our Fucci cell cycle reporters let you label just the nucleus, or both the nucleus and cytoplasm, allowing visualization of cell shape. Although most Fucci probes label only cell nuclei, we offer a truncated version of hGeminin that is able to migrate to the cytoplasm between S and M phases, enabling the morphology of the cell to be visualized.
A Variety of Fucci Probes and Delivery Options
We offer retrovirus- and plasmid-based vectors for the delivery of a variety of Fucci probes. Each probe consists of a red or cyan fluorescent tag, and a deletion mutant of either hCdt1 or hGeminin. Our retroviral vectors can either be transfected as plasmids or transduced as retrovirus in a wide variety of mammalian cells. Once integrated into the host genome, these vectors provide stable, heritable expression of the probes.
The plasmid-based vector, pTRE-CellCycle, allows tightly controlled, tetracycline (Tet)-inducible expression of two Fucci probes from a bidirectional promoter that can be induced using any Tet-On or Tet-Off technology. Expression of these probes causes cell nuclei to turn red during G1 phase and cyan during S, G2, and M phases, allowing complete visual tracking of the cell cycle.