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A.M. Komaromy, V.A. Chiodo, J.J. Alexander, W.W. Hauswirth, G.M. Acland, G.D. Aguirre; Targeting Gene Expression To Canine Cones With The Human Cone Opsin Promoters . Invest. Ophthalmol. Vis. Sci. 2006;47(13):832.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate adeno–associated virus (AAV)–mediated green fluorescent protein (GFP) expression in canine cones using the human red and blue cone opsin promoters.
AAV vectors with serotype 5 capsids containing GFP cDNA under control of either the human red (PR2.1) or blue (HB570) cone opsin promoters were injected into the subretinal space of young, normal dogs. A total of 12 eyes of 6 dogs were injected with 150 µl with 1.2–1.6x1012 vector genomes. Retinal sections were examined for GFP expression after 4 and 8 weeks under fluorescence microscopy. Immunocytochemistry using blue and red/green opsin antibodies helped confirm the targeting specificity of GFP expression.
No adverse effects of the viral vectors or the GFP expression were noted clinically and histologically. The PR2.1 promoter led to specific GFP expression in the red/green cones, evident by GFP fluorescence co–localizing with red/green opsin staining. The entire cells were brightly fluorescent. Alternatively, the HB570 promoter led to the expression of GFP in a subgroup of red/green cones, rods, and in the RPE.
The human cone opsin promoters are effective in directing expression of transferred genes in canine cone photoreceptor cells. While the red cone opsin promoter (PR2.1) showed a high degree of efficiency and specificity in expressing GFP in red/green cones, the blue cone opsin promoter (HB570) was less robust and specific as evidenced by GFP expression in some rod photoreceptors and RPE cells. As red/green cones outnumber the blue cones in mammalian retinas, our results indicate that cone directed gene replacement therapy should be possible with the PR2.1 promoter.
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