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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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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.
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.
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.
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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