title: A neuron-specific gene transfer by a recombinant defective Sindbis virus.
abstract: We examined the possibility that Sindbis virus, an alpha virus with a single-stranded RNA genome, would be applied for neuronal gene transfer. The recombinant defective Sindbis viruses were constructed by replacing the structural genes of Sindbis virus with genes encoding beta-galactosidase (rdSind-lacZ) or enhanced green fluorescent protein (rdSind-EGFP). In neuron-glia cocultures prepared from the neocortex, hippocampus, and striatum, EGFP or beta-galactosidase was expressed selectively in neurons 24 h after infection with rdSind-EGFP or rdSind-lacZ. Most cortical neurons were infected with rdSind-lacZ at a multiplicity of infection (M.O.I.) of 5 while glial cells were little infected. In addition, transient neuron-specific expression of beta-galactosidase was observed near injection sites over the next 3 d following administration of rdSind-lacZ in adult rat. In the cortical neurons infected with rdSind-EGFP, treatment with NMDA induced neuritic blebs and cell body swelling in a Na+-dependent manner. Therefore, recombinant defective Sindbis viruses can be used as an efficient and selective vector for gene transfer into neurons and applied to investigate biological role of target genes delivered into neurons in vitro and in vivo.
authors.affiliation: Department of Pharmacology, Ajou University School of Medicine, Suwon, Kyungkido, South Korea. bjgwag@madang.ajou.ac.kr
authors.forename: B J
authors.initials: BJ
authors.lastname: Gwag
authors.forename: E Y
authors.initials: EY
authors.lastname: Kim
authors.forename: B R
authors.initials: BR
authors.lastname: Ryu
authors.forename: S J
authors.initials: SJ
authors.lastname: Won
authors.forename: H W
authors.initials: HW
authors.lastname: Ko
authors.forename: Y J
authors.initials: YJ
authors.lastname: Oh
authors.forename: Y G
authors.initials: YG
authors.lastname: Cho
authors.forename: S J
authors.initials: SJ
authors.lastname: Ha
authors.forename: Y C
authors.initials: YC
authors.lastname: Sung
mesh_terms: D018354:Alphavirus Infections; D000818:Animals; D002478:Cells, Cultured; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D007763:Lac Operon; D008164:Luminescent Proteins; D051379:Mice; D016202:N-Methylaspartate / Q000633:toxicity; D009457:Neuroglia / Q000166:cytology / Q000821:virology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000821:virology*; D010957:Plasmids; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012845:Sindbis Virus; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D018690:Excitatory Amino Acid Agonists; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D016202:N-Methylaspartate; D001616:beta-Galactosidase