Design & Device

Promoter: 4 copies of NFκB gene

On the basis of NF-κB signaling pathway and its feedback loop dynamics, we provided a device: A high-throughput drug screening system. The system is used to screen targeted drugs in NF-κB signaling pathway and lasts even longer. Natrually, we could estimate whether the kind of drug is good or not.


NF-κB is a DNA binding protein, which means that the dimer could recognize a certain gene sequence and bind with it. Thus, we designed a four coupies of NF-κB gene sequence as the promoter. The downstream gene IκB flag with GFP, which could be a controller in tumor invading，moreover ， easy to observe.

Expriments

4X-NFκB-IκBα-4.20 was tranfected in DU145 cells, Hep-G2, Changliver cells. Since the these kinds of cells have background expression of NF-κB, we use gene knockout technology to remove these parts of gene. These images were taken with a cooled CCD camera 1 day after transfection using the fluorescent microscope. The images all showed that our promoter 4X-NFκB responsed in a good condition by that time, and the difference of brightness of the fluorescent also provide a evidence that our recombainat vector works well. Additionally, we will do some experiments to transfect DU145, 293T and Hela cells with the vector for establishment of stable cell lines for further research.