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Sci Rep. 2016 Jan 27;6:19969. doi: 10.1038/srep19969.

Precision Medicine: Genetic Repair of Retinitis Pigmentosa in Patient-Derived Stem Cells.

Author information

1
Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
2
The Bernard &Shirlee Brown Glaucoma Laboratory, Departments of Ophthalmology, Pathology &Cell Biology, Institute of Human Nutrition, College of Physicians &Surgeons, Columbia University, New York, NY, USA.
3
Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA.

Abstract

Induced pluripotent stem cells (iPSCs) generated from patient fibroblasts could potentially be used as a source of autologous cells for transplantation in retinal disease. Patient-derived iPSCs, however, would still harbor disease-causing mutations. To generate healthy patient-derived cells, mutations might be repaired with new gene-editing technology based on the bacterial system of clustered regularly interspersed short palindromic repeats (CRISPR)/Cas9, thereby yielding grafts that require no patient immunosuppression. We tested whether CRISPR/Cas9 could be used in patient-specific iPSCs to precisely repair an RPGR point mutation that causes X-linked retinitis pigmentosa (XLRP). Fibroblasts cultured from a skin-punch biopsy of an XLRP patient were transduced to produce iPSCs carrying the patient's c.3070G > T mutation. The iPSCs were transduced with CRISPR guide RNAs, Cas9 endonuclease, and a donor homology template. Despite the gene's repetitive and GC-rich sequences, 13% of RPGR gene copies showed mutation correction and conversion to the wild-type allele. This is the first report using CRISPR to correct a pathogenic mutation in iPSCs derived from a patient with photoreceptor degeneration. This important proof-of-concept finding supports the development of personalized iPSC-based transplantation therapies for retinal disease.

PMID:
26814166
PMCID:
PMC4728485
DOI:
10.1038/srep19969
[Indexed for MEDLINE]
Free PMC Article

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