{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T09:10:04Z","timestamp":1790932204197,"version":"4.1.0"},"reference-count":36,"publisher":"eLife Sciences Publications, Ltd","license":[{"start":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T00:00:00Z","timestamp":1623110400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T00:00:00Z","timestamp":1623110400000},"content-version":"am","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T00:00:00Z","timestamp":1623110400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R01EY017950"],"award-info":[{"award-number":["R01EY017950"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001818","name":"Research to Prevent Blindness","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100001818","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["EY017950"],"award-info":[{"award-number":["EY017950"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["elifesciences.org"],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:p>\n                    A missense mutation of collagen type VIII alpha 2 chain (\n                    <jats:italic>COL8A2<\/jats:italic>\n                    ) gene leads to early-onset Fuchs\u2019 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\n                    <jats:italic>COL8A2<\/jats:italic>\n                    expression in corneal endothelial cells, prevented endothelial cell loss, and rescued corneal endothelium pumping function in adult\n                    <jats:italic>Col8a2<\/jats:italic>\n                    mutant mice. There were no adverse sequelae on histology or electroretinography.\n                    <jats:italic>Col8a2<\/jats:italic>\n                    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.\n                  <\/jats:p>","DOI":"10.7554\/elife.55637","type":"journal-article","created":{"date-parts":[[2021,6,8]],"date-time":"2021-06-08T08:00:59Z","timestamp":1623139259000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.55637","source":"Crossref","is-referenced-by-count":53,"title":["Start codon disruption with CRISPR\/Cas9 prevents murine Fuchs\u2019 endothelial corneal dystrophy"],"prefix":"10.7554","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6133-4918","authenticated-orcid":true,"given":"Hironori","family":"Uehara","sequence":"first","affiliation":[{"name":"Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felipe","family":"Pereira","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of Virginia, Charlottesville, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siddharth","family":"Narendran","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of Virginia, Charlottesville, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susie","family":"Choi","sequence":"additional","affiliation":[{"name":"Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sai","family":"Bhuvanagiri","sequence":"additional","affiliation":[{"name":"Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinlu","family":"Liu","sequence":"additional","affiliation":[{"name":"Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sangeetha","family":"Ravi Kumar","sequence":"additional","affiliation":[{"name":"Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Austin","family":"Bohner","sequence":"additional","affiliation":[{"name":"Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lara","family":"Carroll","sequence":"additional","affiliation":[{"name":"Moran Eye Center, Department of Ophthalmology and Visual Sciences, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bonnie","family":"Archer","sequence":"additional","affiliation":[{"name":"Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Zhang","sequence":"additional","affiliation":[{"name":"Division of Epidemiology, Department of Internal Medicine, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Liu","sequence":"additional","affiliation":[{"name":"Division of Epidemiology, Department of Internal Medicine, University of Utah, Salt Lake City, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangping","family":"Gao","sequence":"additional","affiliation":[{"name":"Gene Therapy Center, Department of Microbiology and Physiological Science Systems, University of Massachusetts Medical School, Worcester, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jayakrishna","family":"Ambati","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, University of Virginia, Charlottesville, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert S","family":"Jun","sequence":"additional","affiliation":[{"name":"Wilmer Eye Institute, Johns Hopkins University, Baltimore, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Balamurali K","family":"Ambati","sequence":"additional","affiliation":[{"name":"Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4374","published-online":{"date-parts":[[2021,6,8]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms14898","article-title":"Genome-wide association study identifies three novel loci in fuchs endothelial corneal dystrophy","volume":"8","author":"Afshari","year":"2017","journal-title":"Nature Communications"},{"key":"bib2","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1111\/cge.12191","article-title":"Genetics of the corneal endothelial dystrophies: an evidence-based review","volume":"84","author":"Aldave","year":"2013","journal-title":"Clinical Genetics"},{"key":"bib3","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-017-1363-3","article-title":"Intergenic disease-associated regions are abundant in novel transcripts","volume":"18","author":"Bartonicek","year":"2017","journal-title":"Genome Biology"},{"key":"bib4","doi-asserted-by":"publisher","first-page":"2415","DOI":"10.1093\/hmg\/10.21.2415","article-title":"Missense mutations in COL8A2, the gene encoding the alpha2 chain of type VIII collagen, cause two forms of corneal endothelial dystrophy","volume":"10","author":"Biswas","year":"2001","journal-title":"Human Molecular Genetics"},{"key":"bib5","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1016\/j.exer.2011.06.004","article-title":"Molecular mechanisms underlying the corneal endothelial pump","volume":"95","author":"Bonanno","year":"2012","journal-title":"Experimental Eye Research"},{"key":"bib6","doi-asserted-by":"publisher","first-page":"1520","DOI":"10.1002\/humu.22141","article-title":"The KAT6B-related disorders genitopatellar syndrome and ohdo\/SBBYS syndrome have distinct clinical features reflecting distinct molecular mechanisms","volume":"33","author":"Campeau","year":"2012","journal-title":"Human Mutation"},{"key":"bib7","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using CRISPR\/Cas systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"bib8","doi-asserted-by":"publisher","first-page":"E2253","DOI":"10.1073\/pnas.1616892114","article-title":"Synaptic vesicle glycoprotein 2C (SV2C) modulates dopamine release and is disrupted in Parkinson disease","volume":"114","author":"Dunn","year":"2017","journal-title":"PNAS"},{"key":"bib9","volume-title":"Eye Banking Stastical Report","author":"EBAA","year":"2016"},{"key":"bib10","doi-asserted-by":"publisher","first-page":"1934","DOI":"10.1167\/iovs.04-0937","article-title":"Inheritance of a novel COL8A2 mutation defines a distinct early-onset subtype of fuchs corneal dystrophy","volume":"46","author":"Gottsch","year":"2005","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib11","doi-asserted-by":"publisher","first-page":"1232","DOI":"10.1096\/fj.04-3019com","article-title":"Targeted disruption of Col8a1 and Col8a2 genes in mice leads to anterior segment abnormalities in the eye","volume":"19","author":"Hopfer","year":"2005","journal-title":"The FASEB Journal"},{"key":"bib12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0046742","article-title":"Differing roles for TCF4 and COL8A2 in central corneal thickness and Fuchs endothelial corneal dystrophy","volume":"7","author":"Igo","year":"2012","journal-title":"PLOS ONE"},{"key":"bib13","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1093\/hmg\/ddr473","article-title":"An alpha 2 collagen VIII transgenic knock-in mouse model of fuchs endothelial corneal dystrophy shows early endothelial cell unfolded protein response and apoptosis","volume":"21","author":"Jun","year":"2012","journal-title":"Human Molecular Genetics"},{"key":"bib14","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1038\/nmeth.3284","article-title":"Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells","volume":"12","author":"Kim","year":"2015","journal-title":"Nature Methods"},{"key":"bib15","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1038\/308241a0","article-title":"Point mutations close to the AUG initiator Codon affect the efficiency of translation of rat preproinsulin in vivo","volume":"308","author":"Kozak","year":"1984","journal-title":"Nature"},{"key":"bib16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0183719","article-title":"TGC repeat expansion in the TCF4 gene increases the risk of Fuchs' endothelial corneal dystrophy in Australian cases","volume":"12","author":"Kuot","year":"2017","journal-title":"PLOS ONE"},{"key":"bib17","doi-asserted-by":"publisher","first-page":"1236","DOI":"10.1038\/nprot.2007.135","article-title":"A protocol for rapid generation of recombinant adenoviruses using the AdEasy system","volume":"2","author":"Luo","year":"2007","journal-title":"Nature Protocols"},{"key":"bib18","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1126\/science.1232033","article-title":"RNA-guided human genome engineering via Cas9","volume":"339","author":"Mali","year":"2013","journal-title":"Science"},{"key":"bib19","doi-asserted-by":"publisher","first-page":"6718","DOI":"10.1167\/iovs.12-9669","article-title":"Endothelial Cdkn1a (p21) overexpression and accelerated senescence in a mouse model of fuchs endothelial corneal dystrophy","volume":"53","author":"Matthaei","year":"2012","journal-title":"Investigative Ophthalmology & Visual Science"},{"key":"bib20","doi-asserted-by":"publisher","first-page":"1887","DOI":"10.1167\/iovs.12-11021","article-title":"L450W and Q455K Col8a2 knock-in mouse models of fuchs endothelial corneal dystrophy show distinct phenotypes and evidence for altered autophagy","volume":"54","author":"Meng","year":"2013","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib21","doi-asserted-by":"publisher","first-page":"742","DOI":"10.1001\/jamaophthalmol.2014.352","article-title":"Descemet stripping automated endothelial keratoplasty after failed penetrating keratoplasty: survival, rejection risk, and visual outcome","volume":"132","author":"Mitry","year":"2014","journal-title":"JAMA Ophthalmology"},{"key":"bib22","doi-asserted-by":"publisher","first-page":"2003","DOI":"10.1167\/iovs.14-16222","article-title":"TCF4 triplet repeat expansion and nuclear RNA foci in Fuchs' Endothelial corneal dystrophy","volume":"56","author":"Mootha","year":"2015","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib23","doi-asserted-by":"publisher","first-page":"4865","DOI":"10.1167\/iovs.15-17082","article-title":"Trinucleotide repeat expansion in the TCF4 Gene in Fuchs' Endothelial Corneal Dystrophy in Japanese","volume":"56","author":"Nakano","year":"2015","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib24","doi-asserted-by":"publisher","first-page":"7674","DOI":"10.1167\/iovs.14-15297","article-title":"Genetic association of TCF4 intronic polymorphisms, CTG18.1 and rs17089887, with Fuchs' endothelial corneal dystrophy in an Indian population","volume":"55","author":"Nanda","year":"2014","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib25","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1016\/j.neuron.2017.10.004","article-title":"Virus-Mediated genome editing via Homology-Directed repair in Mitotic and postmitotic cells in mammalian brain","volume":"96","author":"Nishiyama","year":"2017","journal-title":"Neuron"},{"key":"bib26","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1093\/hmg\/10.4.395","article-title":"Fusion of the MORF and CBP genes in acute myeloid leukemia with the t(10;16)(q22;p13)","volume":"10","author":"Panagopoulos","year":"2001","journal-title":"Human Molecular Genetics"},{"key":"bib27","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/j.ajhg.2009.12.001","article-title":"Missense mutations in TCF8 cause late-onset fuchs corneal dystrophy and interact with FCD4 on chromosome 9p","volume":"86","author":"Riazuddin","year":"2010","journal-title":"The American Journal of Human Genetics"},{"key":"bib28","doi-asserted-by":"publisher","first-page":"2128","DOI":"10.1101\/gr.2973604","article-title":"Human ORFeome version 1.1: a platform for reverse proteomics","volume":"14","author":"Rual","year":"2004","journal-title":"Genome Research"},{"key":"bib29","doi-asserted-by":"publisher","first-page":"32","DOI":"10.3109\/13816810.2012.726396","article-title":"Confirmation of the association between the TCF4 risk allele and Fuchs endothelial corneal dystrophy in patients from the Midwestern United States","volume":"34","author":"Stamler","year":"2013","journal-title":"Ophthalmic Genetics"},{"key":"bib30","doi-asserted-by":"publisher","first-page":"246","DOI":"10.1001\/jamaophthalmol.2014.3923","article-title":"Factors associated with corneal graft survival in the cornea donor study","volume":"133","author":"Sugar","year":"2015","journal-title":"JAMA Ophthalmology"},{"key":"bib31","doi-asserted-by":"publisher","first-page":"815","DOI":"10.1136\/bjo.65.12.815","article-title":"Dual staining of corneal endothelium with trypan blue and alizarin red S: importance of pH for the dye-lake reaction","volume":"65","author":"Taylor","year":"1981","journal-title":"British Journal of Ophthalmology"},{"key":"bib32","doi-asserted-by":"publisher","first-page":"321","DOI":"10.2147\/OPTH.S83467","article-title":"Fuchs endothelial corneal dystrophy: current perspectives","volume":"10","author":"Vedana","year":"2016","journal-title":"Clinical Ophthalmology"},{"key":"bib33","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0200005","article-title":"Gene expression in the corneal endothelium of fuchs endothelial corneal dystrophy patients with and without expansion of a trinucleotide repeat in TCF4","volume":"13","author":"Wieben","year":"2018","journal-title":"PLOS ONE"},{"key":"bib34","doi-asserted-by":"publisher","first-page":"3636","DOI":"10.1167\/iovs.19-27689","article-title":"Gene expression and missplicing in the corneal endothelium of patients with a TCF4 trinucleotide repeat expansion without Fuchs' Endothelial corneal dystrophy","volume":"60","author":"Wieben","year":"2019","journal-title":"Investigative Opthalmology & Visual Science"},{"key":"bib35","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms14716","article-title":"Nrl knockdown by AAV-delivered CRISPR\/Cas9 prevents retinal degeneration in mice","volume":"8","author":"Yu","year":"2017","journal-title":"Nature Communications"},{"key":"bib36","doi-asserted-by":"publisher","first-page":"830","DOI":"10.1038\/cr.2017.57","article-title":"Gene and mutation independent therapy via CRISPR-Cas9 mediated cellular reprogramming in rod photoreceptors","volume":"27","author":"Zhu","year":"2017","journal-title":"Cell Research"}],"container-title":["eLife"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/cdn.elifesciences.org\/articles\/55637\/elife-55637-v2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/cdn.elifesciences.org\/articles\/55637\/elife-55637-v2.xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/elifesciences.org\/articles\/55637","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,11]],"date-time":"2023-10-11T20:59:22Z","timestamp":1697057962000},"score":1,"resource":{"primary":{"URL":"https:\/\/elifesciences.org\/articles\/55637"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,8]]},"references-count":36,"alternative-id":["10.7554\/eLife.55637"],"URL":"https:\/\/doi.org\/10.7554\/elife.55637","archive":["CLOCKSS"],"relation":{"is-supplemented-by":[{"id-type":"accession","id":"GSE146999","asserted-by":"subject"}],"has-review":[{"id-type":"doi","id":"10.7554\/eLife.55637.sa2","asserted-by":"object"},{"id-type":"doi","id":"10.7554\/eLife.55637.sa1","asserted-by":"object"}],"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.03.18.996728","asserted-by":"object"}]},"ISSN":["2050-084X"],"issn-type":[{"value":"2050-084X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,8]]},"assertion":[{"value":"single anonymised","name":"peer_review_transparency","label":"Peer review transparency","group":{"name":"peer_review_taxonomy"}},{"value":"other reviewer(s), editor","name":"peer_review_interaction","label":"Peer review interaction","group":{"name":"peer_review_taxonomy"}},{"value":"review summaries, review reports, author\/editor communication, reviewer identities reviewer opt in, editor identities","name":"peer_review_published","label":"Peer review published","group":{"name":"peer_review_taxonomy"}},{"value":"open (sign in with ORCID iD required)","name":"post_publication_commenting","label":"Post publication commenting","group":{"name":"post_publication_commenting"}}],"article-number":"e55637"}}