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Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy
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 the loss of corneal endothelial cells. We demonstrate that CRISPR/Cas9-based postnatal gene editing achieves structural a...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8216720/ https://www.ncbi.nlm.nih.gov/pubmed/34100716 http://dx.doi.org/10.7554/eLife.55637 |
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author | Uehara, Hironori Zhang, Xiaohui Pereira, Felipe Narendran, Siddharth Choi, Susie Bhuvanagiri, Sai Liu, Jinlu Ravi Kumar, Sangeetha Bohner, Austin Carroll, Lara Archer, Bonnie Zhang, Yue Liu, Wei Gao, Guangping Ambati, Jayakrishna Jun, Albert S Ambati, Balamurali K |
author_facet | Uehara, Hironori Zhang, Xiaohui Pereira, Felipe Narendran, Siddharth Choi, Susie Bhuvanagiri, Sai Liu, Jinlu Ravi Kumar, Sangeetha Bohner, Austin Carroll, Lara Archer, Bonnie Zhang, Yue Liu, Wei Gao, Guangping Ambati, Jayakrishna Jun, Albert S Ambati, Balamurali K |
author_sort | Uehara, Hironori |
collection | PubMed |
description | 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 the 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 the 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. |
format | Online Article Text |
id | pubmed-8216720 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-82167202021-06-23 Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy Uehara, Hironori Zhang, Xiaohui Pereira, Felipe Narendran, Siddharth Choi, Susie Bhuvanagiri, Sai Liu, Jinlu Ravi Kumar, Sangeetha Bohner, Austin Carroll, Lara Archer, Bonnie Zhang, Yue Liu, Wei Gao, Guangping Ambati, Jayakrishna Jun, Albert S Ambati, Balamurali K eLife Genetics and Genomics 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 the 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 the 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. eLife Sciences Publications, Ltd 2021-06-08 /pmc/articles/PMC8216720/ /pubmed/34100716 http://dx.doi.org/10.7554/eLife.55637 Text en © 2021, Uehara et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Genetics and Genomics Uehara, Hironori Zhang, Xiaohui Pereira, Felipe Narendran, Siddharth Choi, Susie Bhuvanagiri, Sai Liu, Jinlu Ravi Kumar, Sangeetha Bohner, Austin Carroll, Lara Archer, Bonnie Zhang, Yue Liu, Wei Gao, Guangping Ambati, Jayakrishna Jun, Albert S Ambati, Balamurali K Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title | Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title_full | Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title_fullStr | Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title_full_unstemmed | Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title_short | Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy |
title_sort | start codon disruption with crispr/cas9 prevents murine fuchs’ endothelial corneal dystrophy |
topic | Genetics and Genomics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8216720/ https://www.ncbi.nlm.nih.gov/pubmed/34100716 http://dx.doi.org/10.7554/eLife.55637 |
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