CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa

Wen-Hsuan Wu; Yi-Ting Tsai; Sally Justus; Ting-Ting Lee; Lijuan Zhang; Chyuan-Sheng Lin; Alexander G Bassuk; Vinit B Mahajan; Stephen H Tsang

doi:10.1038/mt.2016.107

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CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa

Journal article

Open access

Peer reviewed

CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa

Wen-Hsuan Wu, Yi-Ting Tsai, Sally Justus, Ting-Ting Lee, Lijuan Zhang, Chyuan-Sheng Lin, Alexander G Bassuk, Vinit B Mahajan and Stephen H Tsang

Molecular therapy, Vol.24(8), pp.1388-1394

08/2016

DOI: 10.1038/mt.2016.107

PMCID: PMC5023380

PMID: 27203441

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Abstract

Massive parallel sequencing enables identification of numerous genetic variants in mutant organisms, but determining pathogenicity of any one mutation can be daunting. The most commonly studied preclinical model of retinitis pigmentosa called the "rodless" (rd1) mouse is homozygous for two mutations: a nonsense point mutation (Y347X) and an intronic insertion of a leukemia virus (Xmv-28). Distinguishing which mutation causes retinal degeneration is still under debate nearly a century after the discovery of this model organism. Here, we performed gene editing using the CRISPR/Cas9 system and demonstrated that the Y347X mutation is the causative variant of disease. Genome editing in the first generation produced animals that were mosaic for the corrected allele but still showed neurofunction preservation despite low repair frequencies. Furthermore, second-generation CRISPR-repaired mice showed an even more robust rescue and amelioration of the disease. This predicts excellent outcomes for gene editing in diseased human tissue, as Pde6b, the mutated gene in rd1 mice, has an orthologous intron-exon relationship comparable with the human PDE6B gene. Not only do these findings resolve the debate surrounding the source of neurodegeneration in the rd1 model, but they also provide the first example of homology-directed recombination-mediated gene correction in the visual system.

DNA Repair

Mutation

Electroretinography

Cyclic Nucleotide Phosphodiesterases, Type 6 - metabolism

Exons

Retinal Degeneration - genetics

Retinitis Pigmentosa - genetics

Homologous Recombination

Mice, Transgenic

Cyclic Nucleotide Phosphodiesterases, Type 6 - genetics

Genetic Loci

Gene Editing

Animals

CRISPR-Cas Systems

Clustered Regularly Interspaced Short Palindromic Repeats

Mice

RNA, Guide

Photoreceptor Cells, Vertebrate - metabolism

Retinal Degeneration - pathology

Photoreceptor Cells, Vertebrate - pathology

Disease Models, Animal

Details

Title: Subtitle: CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa
Creators: Wen-Hsuan Wu - Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, New York, USA
Yi-Ting Tsai - Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, New York, USA
Sally Justus - Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, New York, USA
Ting-Ting Lee - Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, New York, USA
Lijuan Zhang - Shanxi Eye Hospital, Shanxi Medical University, Taiyuan, Shanxi, China
Chyuan-Sheng Lin - Department of Pathology & Cell Biology, Columbia University Medical Center, New York, New York, USA
Alexander G Bassuk - Department of Pediatrics and Neurology, University of Iowa, Iowa City, Iowa, USA
Vinit B Mahajan - Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, Iowa, USA
Stephen H Tsang - Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, New York, New York, USA
Resource Type: Journal article
Publication Details: Molecular therapy, Vol.24(8), pp.1388-1394
DOI: 10.1038/mt.2016.107
PMID: 27203441
PMCID: PMC5023380
NLM abbreviation: Mol Ther
ISSN: 1525-0016
eISSN: 1525-0024
Publisher: United States
Grant note: R01 EY025225 / NEI NIH HHS R01 EY018213 / NEI NIH HHS R01 EY026682 / NEI NIH HHS R01 EY024698 / NEI NIH HHS
Language: English
Date published: 08/2016
Academic Unit: Neurology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Neurology (Pediatrics)
Record Identifier: 9984020652802771

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