title: Targeting gene expression to cones with human cone opsin promoters in recombinant AAV.
abstract: Specific cone-directed therapy is of high priority in the treatment of human hereditary retinal diseases. However, not much information exists about the specific targeting of photoreceptor subclasses. Three versions of the human red cone opsin promoter (PR0.5, 3LCR-PR0.5 and PR2.1), and the human blue cone opsin promoter HB569, were evaluated for their specificity and robustness in targeting green fluorescent protein (GFP) gene expression to subclasses of cones in the canine retina when used in recombinant adeno-associated viral vectors of serotype 5. The vectors were administered by subretinal injection. The promoter PR2.1 led to most effective and specific expression of GFP in the long- and medium-wavelength-absorbing cones (L/M cones) of normal and diseased retinas. The PR0.5 promoter was not effective. Adding three copies of the 35-bp LCR in front of PR0.5 lead to weak GFP expression in L/M cones. The HB569 promoter was not specific, and GFP was expressed in a few L/M cones, some rods and the retinal pigment epithelium. These results suggest that L/M cones, the predominant class of cone photoreceptors in the retinas of dogs and most mammalian species can be successfully targeted using the human red cone opsin promoter.
authors.affiliation: Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA. komaromy@vet.upenn.edu
authors.forename: A M
authors.initials: AM
authors.lastname: Komáromy
authors.forename: J J
authors.initials: JJ
authors.lastname: Alexander
authors.forename: A E
authors.initials: AE
authors.lastname: Cooper
authors.forename: V A
authors.initials: VA
authors.lastname: Chiodo
authors.forename: L G
authors.initials: LG
authors.lastname: Glushakova
authors.forename: G M
authors.initials: GM
authors.lastname: Acland
authors.forename: W W
authors.initials: WW
authors.lastname: Hauswirth
authors.forename: G D
authors.initials: GD
authors.lastname: Aguirre
mesh_terms: D000818:Animals; D003117:Color Vision Defects / Q000378:metabolism / Q000628:therapy; D000229:Dependovirus / Q000235:genetics; D004285:Dogs; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007267:Injections; D023421:Models, Animal; D011401:Promoter Regions, Genetic; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism*; D017299:Rod Opsins / Q000235:genetics*; D052416:Targeted Gene Repair; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes
chemical_list: D017299:Rod Opsins; C095855:long-wavelength opsin; C095853:short-wavelength opsin; D049452:Green Fluorescent Proteins