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H Conrath, G Duisit, S Saleun, Y Cherel, P Moullier, F Rolling; SIV-mediated Gene Transfer Into Rat Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4608.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To evaluate lentiviral vectors mediated rat retinal transduction using SIV pseudotyped with envelope proteins from vesicular stomatitis virus (VSV-G), lyssavirus (MK-G) or 4070A amphotropic MLV. The three pseudotyped lentivirus vectors carried CMV-driven Green Fluorescent Protein (GFP) or ß-Galactosidase (ß-Gal) reporter genes. Methods: Intravitreal and subretinal injections of recombinant SIV were performed in cohorts of wistar rats. GFP and ß-Gal expression were respectively evaluated by direct in vivo retinal imaging and X-Gal staining on paraffin sections. Since VSV-G-pseudotyped SIV vectors were concentrated before use, possible pseudotranduction was evaluated by producing empty SIV particles in the presence of a CMV-GFP plasmid lacking the encapsidation signal. Results: No transgene expression was detected after intravitreal injection of SIV vectors. However, selective transduction of RPE cells was repeatedly obtained after transscleral-transchoroïdal subretinal delivery of VSV-G, MK-G and 4070A pseudotyped SIV. GFP expression was maximum as soon as 2-3 days post administration and up to 5 months with no evidence for pseudotransduction. Conclusions:VSVG, lyssavirus, and 4070A amphotropic pseudotyped lentiviral vectors specifically target RPE cells with early onset of transgene expression. This vector holds promise as gene therapy vector for retinal degenerative disease originating in RPE cells.
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