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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2021
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.
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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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