We are developing novel technologies for imaging signaling pathway activation for high throughput genetic screens. This will consist of luciferase and green fluorescent protein-based DNA construct that will enable the measurement of protein phosphorylation through Bi-Molecular or Intra-Molecular Fluorescence or Luminescence Complementation in cell-based assays. As a proof of principle, we are currently developing constructs to measure PI 3-kinase pathway activation via BAD and FOXO3 phosphorylation and DNA damage signaling via H2AX and CHK2 phosphorylation. These will be cloned into lentiviral vectors which will be used to transduce primary glioma cells, then the cells will be used with whole genome shRNA and chemical library screens.
