A missense mutation of collagen type VIII alpha 2 chain (COL8A2) gene leads to early onset Fuchs' endothelial corneal dystrophy (FECD), which progressively impairs vision through loss of corneal endothelial cells. We demonstrate that CRISPR/Cas9-based postnatal gene editing achieves structural and functional rescue in a mouse model of FECD. A single intraocular injection of an adenovirus encoding both the Cas9 gene and guide RNA (Ad-Cas9-Col8a2gRNA), efficiently knocked down mutant COL8A2 expression in corneal endothelial cells, prevented endothelial cell loss, and rescued corneal endothelium pumping function in adult Col8a2 mutant mice. There were no adverse sequelae on histology or electroretinography. Col8a2 start codon disruption represents a non-surgical strategy to prevent vision loss in early-onset FECD. As this demonstrates the ability of Ad-Cas9-gRNA to restore phenotype in adult post-mitotic cells, this method may be widely applicable to adult-onset diseases, even in tissues affected with disorders of non-reproducing cells.
High-throughput Sequencing data have been deposited in GEO under accession codes GSE146999. Source data files have been provided as excel files.
Start codon disruption with CRISPR/Cas9 prevents murine Fuchs' endothelial corneal dystrophyNCBI Gene Expression Omnibus, GSE146999.
- Balamurali K Ambati
- Balamurali K Ambati
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Animal experimentation: This study was conducted in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#15-11024 and #18-10016) of the University of Utah.
- Constance L Cepko, Harvard Medical School, United States
- Received: January 31, 2020
- Accepted: June 7, 2021
- Accepted Manuscript published: June 8, 2021 (version 1)
© 2021, Uehara et al.
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