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J.N. Mallam, M.Y. Hurwitz, P. Chevez-Barrios, R.L. Hurwitz; Efficient Gene Transfer into Retinal Cells by Modified Ad5/F35 Chimeric Adenovirus Vectors . Invest. Ophthalmol. Vis. Sci. 2003;44(13):443.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: In vitro and in vivo transduction efficiency of human retinoblastoma cells and murine retina using an adenoviral serotype 5 vector containing the fiber domain derived from adenovirus serotype 35 and the gene encoding green fluorescent protein (Ad5/F35-GFP) was compared to an Ad5-GFP vector. Methods: Y79 human retinoblastoma cells were incubated with either viral vector and the expression of the reporter protein was compared using quantitative fluorescence and fluorescent activated cell sorting. Mice were given a single sub-retinal injection of either Ad5-GFP or Ad5/F35-GFP. Eyes were enucleated at various times after injection for histopathologic examination. Results: Both vectors efficiently transduced the Y79 human retinoblastoma cells in vitro, however, the amount of the transgene expressed using Ad5/F35-GFP was more than six fold greater than that when Ad5-GFP was used. In vivo, Ad5/F35-GFP at doses as low as 105 iu efficiently transduced cells in all layers of the retina including photoreceptors, neuronal cells, and müller cells as well as retinal pigment epithelial cells while Ad5-GFP efficiently transduced only retinal pigment epithelia cells. GFP expression persists at least one month after injection. Conclusion: Ad5/F35 chimeric vectors may be superior to Ad5 for gene therapy applications targeting the retina.
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