June 2020
Volume 61, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2020
scAAV.7m8 and VMD2 Promoter as Novel Gene Delivery Tools Targeting Chicken RPE
Author Affiliations & Notes
  • Yan Zhang
    School of Optometry, University of California, Berkeley, Berkeley, California, United States
  • Mei Li
    School of Optometry, University of California, Berkeley, Berkeley, California, United States
    Vision Science Core Gene Delivery Module, University of California, Berkeley, Berkeley, California, United States
  • Alexander Ly
    School of Optometry, University of California, Berkeley, Berkeley, California, United States
    Department of Molecular & Cell Biology, University of California, Berkeley, Berkeley, California, United States
  • John Gerard Flannery
    School of Optometry, University of California, Berkeley, Berkeley, California, United States
    Department of Molecular & Cell Biology, University of California, Berkeley, Berkeley, California, United States
  • Christine Wildsoet
    School of Optometry, University of California, Berkeley, Berkeley, California, United States
  • Footnotes
    Commercial Relationships   Yan Zhang, None; Mei Li, None; Alexander Ly, None; John Flannery, None; Christine Wildsoet, None
  • Footnotes
    Support  NIH Grant R21EY029107, K12EY017269, K08EY023609, R01EY012392, and P30EY003176.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 855. doi:
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    • Get Citation

      Yan Zhang, Mei Li, Alexander Ly, John Gerard Flannery, Christine Wildsoet; scAAV.7m8 and VMD2 Promoter as Novel Gene Delivery Tools Targeting Chicken RPE. Invest. Ophthalmol. Vis. Sci. 2020;61(7):855.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : This study investigated the feasibility and specificity of novel gene delivery tools targeting chicken RPE (retinal pigment epithelium) using a new AAV serotype 7m8 and an RPE-specific promoter VMD2.

Methods : Cultured RPE cells and scleral fibroblasts were obtained from 3-day-old chickens and cultured up to 6 weeks. RPE cells were infected with either scAAV.7m8-VMD2-eGFP or scAAV.7m8-CAG-eGFP at concentration of 1 x 1010 vg/ml after they were confluent. eGFP fluorescence in cultured RPE cells was monitored with a Zeiss LSM 710 Confocal Laser Scanning Microscope every week, for up to 4 weeks. Scleral fibroblasts were also infected with these two viral vectors at the same pattern and observed for up to 2 weeks after administration of viral vectors.

Results : Cultured chicken RPE cells, but not scleral fibroblasts, were successfully transduced with scAAV.7m8-VMD2-eGFP, as indicated by eGFP fluorescence. Specifically, eGFP fluorescence was observed in RPE cells one week after transduction. Its intensity increased thereafter and was sustained throughout the 4-weeks observation period, indicating AAV serotype 7m8 and VMD2 promoter combination specifically transduced chicken RPE cells. On the other hand, both RPE and scleral fibroblasts were successfully transduced with scAAV.7m8-CAG-eGFP, as recorded by strong eGFP fluorescence three days after transduction, suggesting that AAV serotype 7m8 nonselectively infects chicken cells.

Conclusions : The results for scAAV.7m8-VMD2 showing selective transduction of the chicken RPE in vitro suggest that it may serve as a tool for gene delivery targeting RPE. They also open up the possibility of exploring gene therapy in an animal model for myopia as a novel approach to myopia control.

This is a 2020 ARVO Annual Meeting abstract.

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