pTRE-CellCycle Vector

Cell-cycle progression as visualized with Fucci probes

Cell-cycle progression as visualized with Fucci probes. Successive rounds of division in HEK 293 Tet-On Advanced cells were monitored using Fucci probes expressed from the pTRE-CellCycle vector in the presence of doxycycline (Dox). As the cells progressed through the cell cycle, their nuclei varied in color: cells lacking color were transitioning between phases M and G1. The nuclei of cells in G1 phase were then labeled with the red fluorescent protein mCherry. As the cells transitioned between phases G1 and S, they appeared light pink/violet. Finally, the nuclei of cells in S through M phases were labeled with the cyan fluorescent protein AmCyan1.

Retroviral delivery is available for a variety of cell-cycle Fucci probes

Retroviral delivery is available for a variety of cell-cycle Fucci probes. Fucci probes containing Geminin-fluorescent protein (fp) fusions are visible in phases S through M, whereas the probe containing Cdt1-fp (expressed by pRetroX-G1-Red) is visible during the G1 phase of the cell cycle. pRetroX-SG2Mcyto-Red expresses a truncated version of geminin that allows the cell shape to be visualized from phases S through M.

Movie of cell-cycle progression, visualized with Fucci probes

Movie of cell-cycle progression, visualized with Fucci probes. No color: cells transitioning between phases M and G1. Red: cells in G1 phase. Light pink/violet: cells transitioning between phases G1 and S. Cyan: the nuclei of cells in phases S through M.

Tightly controlled, simultaneous expression of two Fucci probes allows complete visual tracking of the cell cycle

Tightly controlled, simultaneous expression of two Fucci probes allows complete visual tracking of the cell cycle. pTRE-CellCycle is a bidirectional, tetracycline (Tet)-inducible expression vector that lets you inducibly express Fucci probes during all phases of the cell cycle (Panel A). In cells transfected with the vector, the Cdt1 fusion is visible through G1 phase, while the Geminin fusion is visible from S through M phases (Panel B). A fluorescence micrograph of HEK 293 Tet-On Advanced cells transfected with the pTRE-CellCycle vector and cultured in the presence of Dox is shown in Panel C.

Retrovirus- and plasmid-based vectors for the delivery of a variety of Fucci probes to monitor the cell cycle

Retrovirus- and plasmid-based vectors for the delivery of a variety of Fucci probes to monitor the cell cycle. Each probe consists of a red or cyan fluorescent tag, and a deletion mutant of either hCdt1 or hGeminin.