1. Protein Engineering/Sythetic Biology projects One of the main directions of my lab is development of novel methods for regulation protein function in living cells. We utilize protein engineering to achieve specific control of protein activity using cell permeable compounds or light. We have several rotation projects focused on development of light-regulated enzymes and non-enzymatic proteins that control signaling. Participation in these projects will expose rotating students to current protein engineering strategies, different methods used for biochemical analysis of engineered protein function, and live cell imaging analysis approaches that utilize epifluorescence, TIRF and spinning disk confocal microscopy.
2. Regulation of endothelial cell-cell interactions and angiogenesis. My lab is interested in defining processes that regulate cell-cell interactions and cells migration. We have rotation projects that focus on the role of Src family kinases in regulation of endothelial adherens junctions and angiogenesis. Using synthetic biology tools, we were able to uncover an unexpected function of Src kinase in regulation of endothelial cell-cell junctions. We also discovered a new critical player that regulates the initial step of angiogenesis, degradation of extracellular matrix. Rotation projects in these two directions will focus on interrogation of specific signaling pathways we identified in our previous studies. The rotations will expose students to different approaches for dissection of signaling pathways, experiments with primary endothelial cells, different imaging methods using epifluorescence, TIRF, confocal, and superresolution microscopy, application of already developed synthetic biology tools for manipulation of protein function in living cells.
3. Development of 3D in vitro blood vessel models. One of our new goals is to establish a cell culture model that will allow us to create a perfused blood vessel in vitro. This model will be employed to study function of endothelial cells in a setup closely mimicking physiological environment. One of the goals is to study interaction of cancer cells with vasculature. This model will enable live imaging of artificial blood vessels and tumor organoids in 3D and manipulation of cell function using synthetic biology tools.
– Mimicking transient activation of protein kinases in living cells .
– Time-Variant SRC Kinase Activation Determines Endothelial Permeability Response.