Research will be directed towards developing a herpes simplex virus vector that can be used for gene transfer into neurons. Model genes to be transferred are E. coli lac Z encoding the histochemical marker enzyme - beta-galactosidase, and mouse BNGF encoding the peptide growth factor - beta-nerve growth factor (NGF). The immediate aims are to optimize expression of transferred genes, to evaluate whether these genes will be expressed when the vector is in a latent state in neurons, and to analyze expression of these genes in the rat central nervous system. Vector constructs will be made including various promoter and regulatory elements. Expression will be evaluated in continuous cell lines and primary neural cultures, as well as in sections of rat brain. Beta-galactosidase will be assessed by in situ and solution assays of enzyme activity, as well as by immunoassay; beta-NGF by bioassays, radioimmunoassays and immune precipitation and gel electrophoresis. Virus stocks of vectors will be assessed by restriction mapping and Southern blots; mRNA transcribed from vectors by Northern blots, S1 analysis and in situ hydrization histochemistry. Herpes vectors appear ideally suited for gene transfer into adult neurons in localized regions of the nervous system. This type of gene transfer has potential applications in the treatment of diseases such as diabetes insipidus, the Lesch-Nyhan syndrome, Parkinson disease, Huntington disease and Alzheimer disease.
| Geller, A I (1991) A system, using neural cell lines, to characterize HSV-1 vectors containing genes which affect neuronal physiology, or neuronal promoters. J Neurosci Methods 36:91-103 |
| Geller, A I; Freese, A (1990) Infection of cultured central nervous system neurons with a defective herpes simplex virus 1 vector results in stable expression of Escherichia coli beta-galactosidase. Proc Natl Acad Sci U S A 87:1149-53 |
