Evaluation of RNA silencing plasmids for Notch signalling modulation
Notch signalling is a direct cell-cell communication pathway that plays a key role in cardiovascular development. Specific removal of different Notch receptors and ligands has shown that they are essential for the formation of a mature vasculature. Mutations in Notch related proteins can lead to different cardiovascular diseases like CADASIL, Alagille syndrome or calcific aortic valve disease. Moreover in animal studies it was shown that deletion of several ligands lead to embryonic lethality, highlighting its prominent role in development.
Our group is interested in the role of Notch signalling in vascular organization with a specific focus on mechanobiology. To understand this relationship we study endothelial cells exposed to shear stress, smooth muscle cells exposed stretch or the interaction between these cell types in response to mechanics. We have found that different components of the Notch pathway respond differently to shear stress or strain. By specifically modulating these proteins and regulators of the pathway, we can gain a mechanistic insight of the cellular processes involved in cardiovascular diseases and tissue engineering
One of the methods to study the role of a protein is silencing through RNA interference. With silencing one blocks the production of a protein by degradation of the specific messenger RNA. In this project we will study the modulation of different proteins that are involved in Notch signalling. For each protein, several inhibitions will be analysed for their effectiveness. This will be done with both western blots and fluorescence microscopy.