Journal article
CRISPR-Cas9-Mediated Correction of the 1.02 kb Common Deletion in CLN3 in Induced Pluripotent Stem Cells from Patients with Batten Disease
CRISPR journal, Vol.1(1), pp.75-87
02/01/2018
DOI: 10.1089/crispr.2017.0015
PMCID: PMC6319325
PMID: 31021193
Abstract
Juvenile neuronal ceroid lipofuscinosis (Batten disease) is a rare progressive neurodegenerative disorder caused by mutations in
CLN3
. Patients present with early-onset retinal degeneration, followed by epilepsy, progressive motor deficits, cognitive decline, and premature death. Approximately 85% of individuals with Batten disease harbor at least one allele containing a 1.02 kb genomic deletion spanning exons 7 and 8. This study demonstrates CRISPR-Cas9-based homology-dependent repair of this mutation in induced pluripotent stem cells generated from two independent patients: one homozygous and one compound heterozygous for the 1.02 kb deletion. Our strategy included delivery of a construct that carried >3 kb of DNA: wild-type
CLN3
sequence and a LoxP-flanked, puromycin resistance cassette for positive selection. This strategy resulted in correction at the genomic DNA and mRNA levels in the two independent patient lines. These CRISPR-corrected isogenic cell lines will be a valuable tool for disease modeling and autologous retinal cell replacement.
Details
- Title: Subtitle
- CRISPR-Cas9-Mediated Correction of the 1.02 kb Common Deletion in CLN3 in Induced Pluripotent Stem Cells from Patients with Batten Disease
- Creators
- Erin R Burnight - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaLaura R Bohrer - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaJoseph C Giacalone - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaDarcey L Klaahsen - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaHeather T Daggett - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaJade S East - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaRobert A Madumba - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaKristan S Worthington - 3Department of Biomedical Engineering, University of Iowa, Iowa City, IowaRobert F Mullins - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaEdwin M Stone - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaBudd A Tucker - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaLuke A Wiley - 2Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa
- Resource Type
- Journal article
- Publication Details
- CRISPR journal, Vol.1(1), pp.75-87
- DOI
- 10.1089/crispr.2017.0015
- PMID
- 31021193
- PMCID
- PMC6319325
- NLM abbreviation
- CRISPR J
- ISSN
- 2573-1599
- eISSN
- 2573-1602
- Publisher
- Mary Ann Liebert, Inc
- Language
- English
- Date published
- 02/01/2018
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; Chemical and Biochemical Engineering; Ophthalmology and Visual Sciences
- Record Identifier
- 9984064566802771
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