Purchase this article with an account.
LG Glushakova, Q Li, AM Timmers, TJ Doyle, V Chiodo, WW Hauswirth; AAV-delivered Human Blue Cone Opsin Promoter Drives the Expression of GFP Reporter Gene Primarily in Rat Cone Photoreceptors . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1452.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To evaluate whether the proximal promoter sequence of the human blue cone opsin gene can target specific gene expression in the rodent retina via rAAV vectors in order to develop an efficient S-cone specific gene delivery system. Methods: A 570 bp upstream sequence of human blue cone opsin gene (BH570) was amplified from human blood. It was then cloned into an rAAV cassette, packaged into viral particles and injected into the subretinal space of male Sprague-Dawley rats at PN 40-48. After 1-3 months post-injection, animals were sacrificed and patterns of GFP reporter gene elaboration analyzed in photoreceptor cells (PR's) by immunohistochemistry. Results: Subretinal injections of rAAV.BH570.GFP resulted in photoreceptor-exclusive GFP located predominantly in the central and inferior retina. GFP- and cone-specific antiserum co-immunostaining revealed that GFP was confined mainly to cone photoreceptors: at least 63% ± 10% of the GFP positive PR cells were recognized as cones. The S- vs. M/L-cone specificity of this expression is being currently evaluated with cone subtype-specific antibodies. Conclusion: rAAV delivered 570 bp human blue cone proximal promoter sequence drives GFP reporter gene statement in rat PR 's, predominantly cones. Supported by: EY07864, EY11123, EY11596, NS36302, FFB, MVRF, RPB. CR: P
This PDF is available to Subscribers Only