Correction of NR2E3 Associated Enhanced S-cone Syndrome Patient-specific iPSCs using CRISPR-Cas9

Laura R Bohrer; Luke A Wiley; Erin R Burnight; Jessica A Cooke; Joseph C Giacalone; Kristin R Anfinson; Jeaneen L Andorf; Robert F Mullins; Edwin M Stone; Budd A Tucker

doi:10.3390/genes10040278

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Correction of NR2E3 Associated Enhanced S-cone Syndrome Patient-specific iPSCs using CRISPR-Cas9

Journal article

Open access

Peer reviewed

Correction of NR2E3 Associated Enhanced S-cone Syndrome Patient-specific iPSCs using CRISPR-Cas9

Laura R Bohrer, Luke A Wiley, Erin R Burnight, Jessica A Cooke, Joseph C Giacalone, Kristin R Anfinson, Jeaneen L Andorf, Robert F Mullins, Edwin M Stone and Budd A Tucker

Genes, Vol.10(4), p.278

04/05/2019

DOI: 10.3390/genes10040278

PMCID: PMC6523438

PMID: 30959774

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https://doi.org/10.3390/genes10040278View

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Abstract

Enhanced S-cone syndrome (ESCS) is caused by recessive mutations in the photoreceptor cell transcription factor . Loss of is characterized by repression of rod photoreceptor cell gene expression, over-expansion of the S-cone photoreceptor cell population, and varying degrees of M- and L-cone photoreceptor cell development. In this study, we developed a CRISPR-based homology-directed repair strategy and corrected two different disease-causing mutations in patient-derived induced pluripotent stem cells (iPSCs) generated from two affected individuals. In addition, one patient's iPSCs were differentiated into retinal cells and transcription was evaluated in CRISPR corrected and uncorrected clones. The patient's c.119-2A>C mutation caused the inclusion of a portion of intron 1, the creation of a frame shift, and generation of a premature stop codon. In summary, we used a single set of CRISPR reagents to correct different mutations in iPSCs generated from two individuals with ESCS. In doing so we demonstrate the advantage of using retinal cells derived from affected patients over artificial in vitro model systems when attempting to demonstrate pathophysiologic mechanisms of specific mutations.

Genetic Therapy

Genetic Predisposition to Disease

Eye Diseases, Hereditary - pathology

Orphan Nuclear Receptors - therapeutic use

Retinal Degeneration - genetics

Humans

Induced Pluripotent Stem Cells - transplantation

Gene Expression Regulation, Developmental - genetics

Retinal Rod Photoreceptor Cells - metabolism

Vision Disorders - pathology

CRISPR-Cas Systems - genetics

Orphan Nuclear Receptors - genetics

Eye Diseases, Hereditary - genetics

Vision Disorders - therapy

Cell Differentiation - genetics

Vision Disorders - genetics

Retinal Degeneration - therapy

Eye Diseases, Hereditary - therapy

Mutation

Retinal Degeneration - pathology

Codon, Nonsense - genetics

Retinal Rod Photoreceptor Cells - pathology

Induced Pluripotent Stem Cells - metabolism

Details

Title: Subtitle: Correction of NR2E3 Associated Enhanced S-cone Syndrome Patient-specific iPSCs using CRISPR-Cas9
Creators: Laura R Bohrer - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. laura-bohrer@uiowa.edu
Luke A Wiley - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. luke-wiley@uiowa.edu
Erin R Burnight - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. erin-burnight@uiowa.edu
Jessica A Cooke - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. jessica-cooke@uiowa.edu
Joseph C Giacalone - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. joseph-giacalone@uiowa.edu
Kristin R Anfinson - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. kristin-anfinson@uiowa.edu
Jeaneen L Andorf - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. jeaneen-andorf@uiowa.edu
Robert F Mullins - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. robert-mullins@uiowa.edu
Edwin M Stone - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. edwin-stone@uiowa.edu
Budd A Tucker - Institute for Vision Research, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52241, USA. budd-tucker@uiowa.edu
Resource Type: Journal article
Publication Details: Genes, Vol.10(4), p.278
DOI: 10.3390/genes10040278
PMID: 30959774
PMCID: PMC6523438
NLM abbreviation: Genes (Basel)
ISSN: 2073-4425
eISSN: 2073-4425
Publisher: Switzerland
Grant note: P30 EY025580 / NEI NIH HHS R01 EY026008 / NEI NIH HHS RO1-EY026008 / NEI NIH HHS T32 GM007337 / NIGMS NIH HHS
Language: English
Date published: 04/05/2019
Academic Unit: The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Ophthalmology and Visual Sciences
Record Identifier: 9984070494802771

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