Fluorescent visualization of cancer cellsby monitoring cellular polyamines

我们正在开发一种结合AZ的多胺依赖移码机制、内源性细胞多胺传感器和荧光蛋白技术的新方法来可视化癌细胞。我们利用AZ1 mRNA的移码区和两个荧光蛋白（ECFP和Keima-Red）构建了一个多胺传感器结构。然而，当构建物转染到细胞中时，没有观察到多胺依赖性的Keima-Red荧光增加。因此，我们利用AZ1 mRNA的整个蛋白质编码区，将EGFP基因插入假结结构的下游，改进了传感器结构。转染该结构的细胞显示出多胺依赖性的EGFP荧光和移码产物的增加。继续进一步分析细胞中的多胺反应。

We are developing a novel method to visualize cancer cells by combining the polyamine-dependent frameshift mechanism of AZ, an endogenous cellular polyamine sensor, and the fluorescent protein techniques. We created a polyamine sensor construct with frameshift region of AZ1 mRNA flanked by two fluorescent proteins (ECFP and Keima-Red). When the construct was transfected into the cells, however, polyamine-dependent increase of Keima-Red fluorescence was not observed. Therefore, we improved the sensor construct using the entire protein coding region of AZ1 mRNA with EGFP gene that is inserted immediately downstream of the pseudoknot structure. Cells transfected with this construct showedpolyamine-dependent increase of EGFP fluorescence and frameshift product. Further analysis of polyamine response in the cells is continued.