Journal article
Using CRISPR-Cas9 to Generate Gene-Corrected Autologous iPSCs for the Treatment of Inherited Retinal Degeneration
Molecular Therapy, Vol.25(9), pp.1999-2013
09/06/2017
DOI: 10.1016/j.ymthe.2017.05.015
PMCID: PMC5589061
PMID: 28619647
Abstract
Patient-derived induced pluripotent stem cells (iPSCs) hold great promise for autologous cell replacement. However, for many inherited diseases, treatment will likely require genetic repair pre-transplantation. Genome editing technologies are useful for this application. The purpose of this study was to develop CRISPR-Cas9-mediated genome editing strategies to target and correct the three most common types of disease-causing variants in patient-derived iPSCs: (1) exonic, (2) deep intronic, and (3) dominant gain of function. We developed a homology-directed repair strategy targeting a homozygous Alu insertion in exon 9 of male germ cell-associated kinase (MAK) and demonstrated restoration of the retinal transcript and protein in patient cells. We generated a CRISPR-Cas9-mediated non-homologous end joining (NHEJ) approach to excise a major contributor to Leber congenital amaurosis, the IVS26 cryptic-splice mutation in CEP290, and demonstrated correction of the transcript and protein in patient iPSCs. Lastly, we designed allele-specific CRISPR guides that selectively target the mutant Pro23His rhodopsin (RHO) allele, which, following delivery to both patient iPSCs in vitro and pig retina in vivo, created a frameshift and premature stop that would prevent transcription of the disease-causing variant. The strategies developed in this study will prove useful for correcting a wide range of genetic variants in genes that cause inherited retinal degeneration. Using CRISPR-Cas9 to generate gene-corrected autologous iPSCs for the treatment of inherited retinal degeneration, induced pluripotent stem cells, and genome editing hold great promise for autologous cell replacement therapies for inherited diseases. Here, Burnight and colleagues developed CRISPR-Cas9-mediated genome editing strategies to target and repair the three most common types of retinal disease-causing variants in patient-derived iPSCs: exonic, deep intronic, and dominant gain of function.
Details
- Title: Subtitle
- Using CRISPR-Cas9 to Generate Gene-Corrected Autologous iPSCs for the Treatment of Inherited Retinal Degeneration
- Creators
- Erin R Burnight - University of IowaManav Gupta - Harvard Medical SchoolLuke A Wiley - University of IowaKristin R Anfinson - University of IowaAudrey Tran - Harvard Medical SchoolRobinson Triboulet - Harvard Medical SchoolJeremy M Hoffmann - University of IowaDarcey L Klaahsen - University of IowaJeaneen L Andorf - University of IowaChunhua Jiao - University of IowaElliott H Sohn - University of IowaMalavika K Adur - Department of Animal Sciences, Iowa State University, Ames, IA 50011, USAJason W Ross - Department of Animal Sciences, Iowa State University, Ames, IA 50011, USARobert F Mullins - University of IowaGeorge Q Daley - Harvard Medical SchoolThorsten M Schlaeger - Harvard Medical SchoolEdwin M Stone - University of IowaBudd A Tucker - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Molecular Therapy, Vol.25(9), pp.1999-2013
- DOI
- 10.1016/j.ymthe.2017.05.015
- PMID
- 28619647
- PMCID
- PMC5589061
- NLM abbreviation
- Mol Ther
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Publisher
- Elsevier Inc
- Grant note
- name: Stephen A. Wynn Foundation; DOI: 10.13039/100000053, name: National Eye Institute, award: R01EY024588, R01EY026008; DOI: 10.13039/100009861, name: Grousbeck Family Foundation
- Language
- English
- Date published
- 09/06/2017
- Academic Unit
- The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
- Record Identifier
- 9983979986602771
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