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Catalytic RNA molecules (ribozymes) have been widely used specifically to suppress gene expression. Neuronal nitric oxide synthase (nNOS) is an important molecule involved in normal central nervous system function (e.g. vasodilation, neurotransmission.) and disease (e.g. oxidative stress). This report is an investigation of the hammerhead ribozyme function and potential in the central nervous system using nNOS as a model. Two antisense hammerhead ribozymes, nNOS-RZ1 and nNOS-RZ2, were designed and constructed against nNOS messenger RNA (mRNA). In vitro (cell-free) experiments demonstrated the ability of both ribozymes to cleave nNOS RNA targets. Ribozyme-mediated reduction of the endogenous nNOS mRNA in human TGW-I-nu neuroblastoma cells was demonstrated by plasmid- and adenovirus-mediated transfections. These results may form the basis for studying neuronal gene expression and for designing RNA-directed therapeutic strategies for neurological diseases that involve oxidative stress.

Type

Journal article

Journal

Neurosci Lett

Publication Date

23/08/2002

Volume

329

Pages

81 - 85

Keywords

Adenoviridae, Central Nervous System, Gene Expression Regulation, Enzymologic, Genetic Vectors, Humans, Molecular Sequence Data, Neuroblastoma, Nitric Oxide, Nitric Oxide Synthase, Nitric Oxide Synthase Type I, Nucleic Acid Conformation, Oxidative Stress, Plasmids, RNA, Catalytic, RNA, Messenger, Tumor Cells, Cultured