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).

Project Start
1999-03-01
Project End
2000-02-29
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
2
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Type
DUNS #
053785812
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
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