The vector Design and Production Core will provide recombinant replication-incompetent herpes simplex virus (HSV)-based vectors to transfer and express selected transgenes in neurons in primary culture (Project 2) and in hippocampal neurons prior to and during ischemia in vivo (Project 3). This strategy will be used to directly test the role of selected known and novel genes, identified by their regional expression in ischemic brain (Project 1), in the regulation of ischemic neuronal death. HSV-based vectors will be constructed by homologous recombination of a plasmid-containing the selected gene under the control of the human cytomegalovirus immediate-early promoter (HCMV-IEp) and the lacZ reporter gene-into the thymidine kinase (tk) locus of an ICP4-replication- incompetent HSV vector propagated in complementing E5 cells. The expression of transgene RNA in vitro will be confirmed by Northern blot, and expression of the protein in vitro by Western blot. Gradient purified vector stocks will then be used to transfer and express the transgene product in vitro (Project 2) and in vivo (Project 3).
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| Simon, Roger P; Meller, Robert; Zhou, An et al. (2012) Can genes modify stroke outcome and by what mechanisms? Stroke 43:286-91 |
| Stevens, Susan L; Leung, Philberta Y; Vartanian, Keri B et al. (2011) Multiple preconditioning paradigms converge on interferon regulatory factor-dependent signaling to promote tolerance to ischemic brain injury. J Neurosci 31:8456-63 |
| Stapels, Martha; Piper, Chelsea; Yang, Tao et al. (2010) Polycomb group proteins as epigenetic mediators of neuroprotection in ischemic tolerance. Sci Signal 3:ra15 |
| Marsh, B J; Williams-Karnesky, R L; Stenzel-Poore, M P (2009) Toll-like receptor signaling in endogenous neuroprotection and stroke. Neuroscience 158:1007-20 |
| West, G Alexander; Golshani, Kiarash J; Doyle, Kristian P et al. (2009) A new model of cortical stroke in the rhesus macaque. J Cereb Blood Flow Metab 29:1175-86 |
| Stevens, Susan L; Ciesielski, Thomas M P; Marsh, Brenda J et al. (2008) Toll-like receptor 9: a new target of ischemic preconditioning in the brain. J Cereb Blood Flow Metab 28:1040-7 |
| Doyle, Kristian P; Yang, Tao; Lessov, Nikola S et al. (2008) Nasal administration of osteopontin peptide mimetics confers neuroprotection in stroke. J Cereb Blood Flow Metab 28:1235-48 |
| Khan, Adil A; Mao, Xiao Ou; Banwait, Surita et al. (2008) Regulation of hypoxic neuronal death signaling by neuroglobin. FASEB J 22:1737-47 |
| Pignataro, Giuseppe; Maysami, Samaneh; Studer, Francesca E et al. (2008) Downregulation of hippocampal adenosine kinase after focal ischemia as potential endogenous neuroprotective mechanism. J Cereb Blood Flow Metab 28:17-23 |
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