Cellular dissection of herpes simplex encephalitis with iPS cells

Title: Cellular dissection of herpes simplex encephalitis with iPS cells I. Background Information The proposed research requires implementation of several technical procedures that my lab has substantial experience with and performs on a regular basis. In addition, the work requires handling of herpes simplex virus type 1 (HSV-1) using BSL2 guidelines. I have been studying these herpesviruses for over a decade and have a BSL2 facility dedicated to herpesvirus propagation. II. Services to be Performed 1) Purified stocks of HSV-1 recombinant isolates will be produced on a continuing basis. These stocks include fluorescent reporter variants and mutants lacking ICP0 or deubiquitinase activity. These viruses have already been developed in my laboratory. 2) Neural stem cells will be cultured and differentiated in preparation for infection with HSV-1 stocks. 3) Live-cell fluorescence microscopy assays will be conducted to assess the progress of HSV-1 infection in neurons derived from patient and control samples. This will be carried out in our BSL2 microscopy facility designed specifically for this purpose. 4) Measurements of viral titer, propagation kinetics, cell entry kinetics, and gene expression will be applied to the neural model.