Enhanced systemic transgene expression after nonviral salivary gland transfection using a novel endonuclease inhibitor/DNA formulation
| Autor: | Parker Ea, Michael J. Bennett, Udove Ja, McMahon Bm, Chen Yj, Niedzinski Ej, Olson Dc, Scollay R, Park H |
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| Rok vydání: | 2003 |
| Předmět: |
Male
Transgene Genetic enhancement Genetic Vectors Gene Expression Biology Transfection Polymerase Chain Reaction Salivary Glands Rats Sprague-Dawley chemistry.chemical_compound Endonuclease Plasmid Gene expression Genetics Animals Transgenes Molecular Biology Dose-Response Relationship Drug Aurintricarboxylic Acid Genetic transfer DNA Genetic Therapy Endonucleases Molecular biology Rats Zinc chemistry biology.protein Molecular Medicine Plasmids |
| Zdroj: | Gene Therapy. 10:2133-2138 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3302125 |
| Popis: | Gene transfer to the major salivary glands is an attractive method for the systemic delivery of therapeutic proteins. To date, nonviral gene transfer to these glands has resulted in inadequate systemic protein concentrations. We believe that identification of the barriers responsible for this inefficient transfection will enable the development of enhanced nonviral gene transfer in salivary glands and other tissues. One potential barrier is the degradation of plasmid DNA by endonucleases. To test this hypothesis, we coadministered two endonuclease inhibitors ((zinc and aurintricarboxylic acid (ATA)) with plasmid DNA, containing the secreted alkaline phosphatase gene (SEAP), to the submandibular glands of rats. The effect of zinc and ATA on SEAP expression, tissue accumulation of plasmid DNA, and plasmid DNA stability was then characterized. We observed that mixtures containing zinc/DNA, ATA/DNA, and zinc/ATA/DNA significantly enhanced both systemic transgene expression and the amount of plasmid DNA associated with treated tissues. The relative endonuclease inhibitory activity of zinc, ATA, and zinc/ATA correlated with the observed effects on transfection efficacy. The use of zinc/ATA enhanced the efficacy of salivary gland transfection by at least 1000-fold versus DNA alone. Importantly, this improved performance resulted in robust systemic secretion of an exogenous protein (SEAP), thus demonstrating the potential this nonviral gene transfer technology has as a method to treat systemic protein deficiencies. |
| Databáze: | OpenAIRE |
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