title: Precision Medicine: Genetic Repair of Retinitis Pigmentosa in Patient-Derived Stem Cells.
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.
authors.affiliation: Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Bassuk
authors.affiliation: 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.
authors.forename: Andrew
authors.initials: A
authors.lastname: Zheng
authors.affiliation: 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.
authors.forename: Yao
authors.initials: Y
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Tsang
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA.
authors.forename: Vinit B
authors.initials: VB
authors.lastname: Mahajan
mesh_terms: D064113:CRISPR-Cas Systems; D002478:Cells, Cultured; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics; D000072669:Gene Editing; D023061:Gene Order; D018390:Gene Targeting; D050172:Genes, X-Linked; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008297:Male; D009154:Mutation; D057285:Precision Medicine; D000094704:RNA, Guide, CRISPR-Cas Systems; D012174:Retinitis Pigmentosa / Q000175:diagnosis / Q000235:genetics* / Q000628:therapy; D033581:Stem Cell Transplantation
chemical_list: D005136:Eye Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; C099929:RPGR protein, human
references.citation: Nat Genet. 2000 Aug;25(4):462-6
references.pmid: 10932196
references.citation: N Engl J Med. 2015 May 14;372(20):1887-97
references.pmid: 25938638
references.citation: Nature. 2006 Nov 9;444(7116):203-7
references.pmid: 17093405
references.citation: Cell. 2007 Nov 30;131(5):861-72
references.pmid: 18035408
references.citation: N Engl J Med. 2008 May 22;358(21):2231-9
references.pmid: 18441371
references.citation: Nat Methods. 2009 Nov;6(11):805-8
references.pmid: 19838168
references.citation: Science. 2012 Aug 17;337(6096):816-21
references.pmid: 22745249
references.citation: Nucleic Acids Res. 2013 Oct;41(19):9049-61
references.pmid: 23907390
references.citation: Br J Ophthalmol. 2014 May;98(5):709-11
references.pmid: 24489377
references.citation: Stem Cell Reports. 2014 May 6;2(5):662-74
references.pmid: 24936453
references.citation: Genome Res. 2014 Sep;24(9):1526-33
references.pmid: 25096406
references.citation: JAMA Ophthalmol. 2015 Feb;133(2):219-20
references.pmid: 25375877
references.citation: Lancet. 2015 Feb 7;385(9967):509-16
references.pmid: 25458728
references.citation: Nat Biotechnol. 2015 May;33(5):543-8
references.pmid: 25803306
references.citation: Hum Genet. 2001 Sep;109(3):271-8
references.pmid: 11702207