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EM Surace, A Auricchio, SJ Reich, S Pineles, E O'Connor, JM Wilson, J Bennett; In Utero AAV Vector-Mediated Gene Delivery To The Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4617.
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Purpose:: This study aimed to define the neuron progenitor transduction patterns of adeno-associated virus (AAV) following in utero gene transfer to the developing retina. An AAV serotype 2 genome was used (AAV2) and packed into recombinant AAVs with capsids from other AAV serotypes (2, 5 and 1) to create AAV2/2, 2/5 and 2/1, respectively. Use of the different AAV capsids may alter tropism and other transduction characteristics. Methods:Fetal injections of AAV2/2, 2/5 and 2/1, encoding enhanced green fluorescent protein (EGFP) under a CMV promoter, were performed in pregnant C57Bl/6 and CD1 mice. Vectors were injected into the subretinal space, in order to transduce the proliferative zone of the developing retina on embryonic days 13-14. EGFP expression was evaluated histologically at different time points following injection. Results:Following exo utero gene transfer, mice developed and grew normally. The injected eyes had no macroscopic or microscopic abnormalities. The time of onset of EGFP expression differed between the various AAV vectors. The onset of transgene expression for AAV 2/2, 2/5 and 2/1 was 9 days, 3 days and 24 hours respectively. All AAVs tested efficiently transduced retinal pigment epithelium (RPE) cells. AAV2/1 displayed a high tropism for retinal progenitors. On postnatal day 0 (P0), EGFP expression was widespread, throughout the entire thickness of the developing retina. On days P14 and P21, expression was more restricted to the outer nuclear layer and in particular to cones. Retinal transduction, although less efficient than with AAV2/1, was observed following in utero delivery of AAV2/2 and 2/5 as well. Conclusion:AAV based-vectors efficiently transduce retinal cell progenitors, thus providing delivery vehicles relevant to the treatment of retinal congenital diseases. In addition, this study provides the first example of AAV-mediated transduction of neuron progenitors following direct delivery of virus to the developing retina in utero. Finally, in utero gene transfer may serve as an alternative to conventional transgenic techniques and provide a means of generating animal models of retinal disease.
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