title: Correction of ornithine-delta-aminotransferase deficiency in a Chinese hamster ovary cell line mediated by retrovirus gene transfer.
abstract: Gyrate atrophy (GA) of the choroid and retina is an autosomal recessive chorioretinal degeneration, caused by deficiency of the mitochondrial matrix enzyme ornithine-delta-aminotransferase (OAT). This deficiency results in the accumulation of ornithine in the body fluids and leads to hyperornithinemia. Although the clinical phenotype is largely confined to the eye, OAT deficiency is a systemic disorder. With the final goal of applying gene therapy to this human genetic disease, we have established an in vitro model to test the correction of OAT enzymatic deficiency in mammalian cells, using OAT recombinant retroviruses. We report the construction of several Moloney murine leukemia virus (MoMLV)-based recombinant retrovirus vectors, in which the human OAT cDNA was placed under the transcriptional control of the mouse phosphoglycerate kinase (PGK) promoter or under the enhancer-promoter regulatory element derived from the MoMLV long terminal repeat (LTR). The retrovirus constructs were packaged in the PG13-GALV cell line and used to transduce C9, an OAT deficient cell line derived from Chinese hamster ovary cells (CHO-K1). We show that the recombinant retrovirus transfers the human OAT (hOAT) gene into C9. Expression of the hOAT gene in the transduced C9 deficient cell line exceeded the OAT mRNA level and enzymatic activity of endogenous human fibroblasts.
authors.affiliation: Genetics and Molecular Immunology Section, National Eye Institue, National Institues of Health, Bethesda, MD 20892.
authors.forename: H D
authors.initials: HD
authors.lastname: Lacorazza
authors.forename: M
authors.initials: M
authors.lastname: Jendoubi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002874:Chromosome Mapping; D006224:Cricetinae; D004274:DNA, Recombinant / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D015799:Gyrate Atrophy / Q000628:therapy; D006801:Humans; D008979:Moloney murine leukemia virus / Q000235:genetics*; D009953:Ornithine-Oxo-Acid Transaminase / Q000172:deficiency* / Q000235:genetics*; D014162:Transfection
chemical_list: D004274:DNA, Recombinant; D009953:Ornithine-Oxo-Acid Transaminase