Transduction of foreign regulatory sequences by a replication-defective herpes simplex virus type 1: the rat neuron-specific enolase promoter.
Herpes simplex virus type 1 (HSV-1) can transduce genes into non-proliferating cells such as neurons that are refractory to other means of gene transfer. We have been interested to examine the potential usefulness of HSV-1 as a gene transfer vehicle to analyze neuron-specific regulatory sequences. In this study, we have used a replication-defective HSV-1-based vector deleted for the essential immediate early gene 3 (IE3) to transduce a 1.8 kb promoter fragment from the rat neuron-specific enolase gene (nse) linked to the firefly luciferase reporter gene (luc). It has previously been shown that the same promoter fragment is capable of directing neuron-specific expression of a linked reporter gene in transgenic mice. As an internal control for infection and gene expression, we also inserted the chloramphenicol acetyltransferase (cat) gene driven by the SV40 early promoter/enhancer into the thymidine kinase locus of the same vector. We infected (i) non-neuronal BHK-C13 cells which do not express the endogenous nse gene, (ii) differentiated and non-differentiated pheochromocytoma PC12 cells as well as (iii) N1E-115 neuroblastoma cells, all of which do express endogenous nse. All three cell types produced luciferase upon infection, indicating that the same nse promoter fragment that has previously been shown to be regulated in a cell-specific manner in transgenic mice, was not regulated cell type-specifically in the context of the HSV-1 genome.(ABSTRACT TRUNCATED AT 250 WORDS)
Roemer
K
K
Center for Molecular Genetics, University of California, San Diego, La Jolla 92093, USA.
Johnson
P A
PA
Friedmann
T
T
D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D006224:Cricetinae; D003673:Defective Viruses / Q000235:genetics*; D005786:Gene Expression Regulation; D017781:Genes, Immediate-Early; D017930:Genes, Reporter; D005822:Genetic Vectors; D007668:Kidney; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D009447:Neuroblastoma; D009474:Neurons / Q000201:enzymology; D009928:Organ Specificity; D016716:PC12 Cells; D010751:Phosphopyruvate Hydratase / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D008156:Luciferases; D015500:Chloramphenicol O-Acetyltransferase; D010751:Phosphopyruvate Hydratase