Premo™ FUCCI Cell Cycle Sensor is a fluorescent, two-color sensor of cell cycle progression and division in live cells. It allows accurate and sensitive cell cycle analysis of individual cells or a population of cells by fluorescence microscopy, flow cytometry, or high-content imaging. The Premo™ FUCCI Cell Cycle Sensor is delivered by highly efficient BacMam 2.0 technology, enabling cell cycle studies in essentially any cell type. Provided in a ready-to-use format—simply add, incubate, and image—Premo™ FUCCI affords highly efficient transient expression in cell lines, primary cells, and stem cells.

Premo™ FUCCI Cell Cycle Sensor is: •Accurate:Cell-cycle controlled expression of bright GFP and RFP indicators for live cell analysis of individual cells or populations •Highly Efficient:>90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons •Fast and Convenient:Simply add Premo™ FUCCI Cell Cycle Sensor to your cells in complete medium, incubate overnight, and analyze •Safe:BacMam reagents are non-integrating and non-replicating in mammalian cells, lack observable cytopathic effects, and are suitable for biosafety level (BSL) 1 handling.

Study Cell Cycle in the Relevant Cellular Context If you are studying cell cycle regulators, cell differentiation, or are developing anti-cancer compounds against cell cycle-related targets, Premo™ FUCCI is a superbly sensitive and accurate sensor tool. The efficient transduction and transient expression allows you to conduct these experiments in the biologically relevant cell types. Simple, Quick Cell Cycle Visualization Simply add the ready-to-use Premo™ FUCCI cell cycle sensor to your cells in complete medium, incubate overnight, then visualize cell cycle progression in populations of cells using fluorescence microscopy, flow cytometry, or high content imaging instruments. Cells change from red in the G1 to yellow in the G1/S interphase and green in S, G2, and M phases, as fusions of emGFP and TagRFP coupled to two cell cycle-regulated proteins are expressed and degraded.

Premo™ FUCCI细胞周期传感器的工作原理

Premo™ FUCCI细胞周期传感器是一种基于荧光蛋白的系统，它使用了与不同细胞周期调控因子融合的红色 (RFP) 和绿色 (GFP) 荧光蛋白： cdt1和geminin。

在细胞周期过程中，上述两种蛋白被特定的E3泛素连接酶泛素化，经由蛋白酶体降解。 E3连接酶可对上述两种蛋白的活性进行时序调控，从而在细胞周期过程中实现geminin和cdt1水平的双相周期变化。 在细胞周期的G1期，geminin被降解；因此只有RFP标记的cdt1在细胞核内呈现红色荧光。 在S、G2和M期，cdt1被降解，只有GFP标记的geminin保留，从而使细胞核呈蓝色荧光。

在G1/S转换过程中，当cdt1水平下降且geminin水平增高时，这两种蛋白均存在于细胞内，当绿色和红色图像覆盖时，细胞核发出黄色荧光。 这种从红色-黄色-到绿色的动态颜色变化代表了细胞周期和分裂的进程 。