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Targeting gene expression to cones with human cone opsin promoters in recombinant AAV

Gene Therapy volume 15, pages 1049–1055 (2008)Cite this article

A Corrigendum to this article was published on 08 December 2011

A Corrigendum to this article was published on 01 July 2008

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.

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Acknowledgements

This study was supported by NIH Grants EY06855, EY07132, EY11123, EY13132, K12-EY15398, P30-EY01583, NS36302, Foundation Fighting Blindness, Macular Vision Research Foundation and The ONCE International Price for R&D in Biomedicine and New Technologies for the Blind. We thank Jeremy Nathans (Johns Hopkins University—Howard Hughes Medical Institute) for providing the pR2.1-LacZ plasmid; W Clay Smith (University of Florida) for the GFP antibody; Tom Doyle and Min Ding (University of Florida) for assistance in vector production; T Michael Redmond (National Eye Institute, Bethesda, MD, USA) for the RPE65 antibody; Barbara Zangerl (University of Pennsylvania) for the promoter alignments; Amanda Nickle, Gerri Antonini, Alice Eidsen, Tracy Greiner and the staff of the Retinal Disease Studies Facility at the University of Pennsylvania for their technical support; and Mary Leonard for the figures. WWH and the University of Florida have a financial interest in the use of rAAV therapies, and own equity in a company (AGTC Inc.) that might, in the future, commercialize some aspects of this work. All remaining authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA

    A M Komáromy, A E Cooper & G D Aguirre

  2. Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, USA

    J J Alexander & W W Hauswirth

  3. Department of Ophthalmology, University of Florida, Gainesville, FL

    V A Chiodo & W W Hauswirth

  4. Baker Institute, Cornell University, Ithaca, NY, USA

    G M Acland

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  1. A M Komáromy
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Correspondence to A M Komáromy.

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Komáromy, A., Alexander, J., Cooper, A. et al. Targeting gene expression to cones with human cone opsin promoters in recombinant AAV. Gene Ther 15, 1049–1055 (2008). https://doi.org/10.1038/gt.2008.32

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