September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
CRISPR/dCas9 to treat neovascular age-related macular degeneration
Author Affiliations & Notes
  • Katie L Pennington
    Ophthalmology and Visual Sciences, University of Utah Moran Eye Center, Salt Lake City, Utah, United States
  • Sandeep Kumar
    Department of Ophthalmology, Baylor College of Medicine Cullen Eye Institute, Houston, Texas, United States
  • Alex Jones
    Department of Pathology, University of Utah Huntsman Cancer Institute, Salt Lake City, Utah, United States
  • Margaret M DeAngelis
    Ophthalmology and Visual Sciences, University of Utah Moran Eye Center, Salt Lake City, Utah, United States
  • Yingbin Fu
    Department of Ophthalmology, Baylor College of Medicine Cullen Eye Institute, Houston, Texas, United States
    Ophthalmology and Visual Sciences, University of Utah Moran Eye Center, Salt Lake City, Utah, United States
  • Footnotes
    Commercial Relationships   Katie Pennington, None; Sandeep Kumar, None; Alex Jones, None; Margaret DeAngelis, None; Yingbin Fu, None
  • Footnotes
    Support  This work was supported by the National Institutes of Health National Eye Institute (EY014800), and an Unrestricted Grant from Research to Prevent Blindness, Inc., New York, NY, to the Department of Ophthalmology & Visual Sciences, University of Utah, with additional support awarded to Dr. DeAngelis from The Skaggs Foundation for Research and the Carl Marshall Reeves & Mildred Almen Reeves Foundation, Inc. NIH grant 1R01EY022901 awarded to Dr. Fu also supports this work.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4436. doi:
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    • Get Citation

      Katie L Pennington, Sandeep Kumar, Alex Jones, Margaret M DeAngelis, Yingbin Fu; CRISPR/dCas9 to treat neovascular age-related macular degeneration. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4436.

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

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Abstract

Purpose : To develop a CRISPR/dCas9 system to act as an in vivo therapeutic agent for treatment of AMD in a murine LASER-induced choroidal neovascularization model.

Methods : Short guide RNAs (sgRNAs) were designed to target mVEGF for transcriptional repression. The sgRNAs were tested for repression efficiency using a dual luciferase assay. C57BL/6 mice were subretinally injected with lentivirus encoding dCas9KRAB and/ or sgRNA targeting mVEGF. Injected mice were subjected to photocoagulation to induce choroidal neovascularization (CNV). Treated eyes were harvested for analysis of either lesion size or gene expression.

Results : Transfection of dCas9KRAB and sgRNAs targeting mVEGF efficiently repressed expression of firefly luciferase under the control of the mVEGF promoter + 5’ untranslated region (UTR) in cell culture. Initial results indicate that subretinal injection with dCas9KRAB and mVEGF sgRNA prior to LASER treatment reduces neovascular lesion area.

Conclusions : Targeting mVEGF for transcriptional repression in vivo using the dCas9KRAB-sgRNA system has potential as a therapeutic method for the treatment of AMD.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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