Purchase this article with an account.
W.W. Hauswirth, H. Petrs–Silva, S.–H. Min, J.M. Liu, S. Mani, V.A. Chiodo, M. Ding, R. Linden, S.L. Boye; Self–Complementary AAV Vectors Promote Fast and Efficient Transduction of Mouse Retina . Invest. Ophthalmol. Vis. Sci. 2006;47(13):839.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Our goal is to compare the temporal and cell–type transduction properties of self–complementary adeno–associated virus (scAAV) vectors to those of corresponding standard adeno–associated virus (AAV) vectors when delivered to the mouse retina.
Subretinal and intravitreal injections of type 2 and type 5 scAAV and AAV vectors with matching promoters expressing green fluorescent protein (GFP) were performed in neonate and adult mice. Promoters used in the comparison include cytomegalovirus immediate early promoter (CMV), a 472bp proximal mouse opsin promoter (MOPS) and an abbreviated version of the chimeric CMV–chicken ß–actin promoter (smCBA). GFP expression was evaluated in retinal whole mounts and in sections by both direct GFP fluorescence and by GFP immunohistochemistry.
Overall, both intravitreal and subretinal injections in adult mice using scAAV vectors express their passenger gene much earlier than standard AAV. With scAAV5 MOPS–GFP injected subretinally, expression was seen in photoreceptors as early as 3 days post–injection whereas it took 7–10 days to first detect GFP expressed from an AAV5 vector containing the same promoter and cDNA. With scAAV2–CMV–GFP injected subretinally in adult mice, expression was also significant at 3 days post–injection in both photoreceptors and the RPE whereas it was undetectable for the corresponding AAV2–CMV–GFP vector. Vectors containing the smCBA promoter and expression onset in neonates are also currently being evaluated.
Self–complementary AAV vectors, although unchanged in retinal cell tropism, promote considerably faster transduction of all target retinal cells in the mouse than standard AAV vectors. Therefore scAAV vectored delivery of therapeutic genes may have utility in conditions requiring very rapid passenger gene expression, extending the range of retinal conditions potentially treatable by AAV mediated gene therapy.
This PDF is available to Subscribers Only