{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T10:37:14Z","timestamp":1788345434088,"version":"build-2803163510"},"reference-count":32,"publisher":"Oxford University Press (OUP)","issue":"9","license":[{"start":{"date-parts":[[2021,12,6]],"date-time":"2021-12-06T00:00:00Z","timestamp":1638748800000},"content-version":"vor","delay-in-days":1237,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2018,7,18]],"date-time":"2018-07-18T00:00:00Z","timestamp":1531872000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2018,7,18]],"date-time":"2018-07-18T00:00:00Z","timestamp":1531872000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100004923","name":"Association Fran\u00e7aise contre les Myopathies","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004923","id-type":"DOI","asserted-by":"publisher"}]},{"name":"G\u00eaniris"},{"name":"Agence Nationale pour la Recherche"},{"DOI":"10.13039\/501100002915","name":"Fondation pour la Recherche M\u00e9dicale","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002915","id-type":"DOI","asserted-by":"publisher"}]},{"name":"French Ministry of Research"},{"DOI":"10.13039\/501100004923","name":"AFM-T\u00e9l\u00e9thon","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002915","name":"Fondation pour la Recherche M\u00e9dicale","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002915","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2018,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Heterozygous PAX6 gene mutations leading to haploinsufficiency are the main cause of congenital aniridia, a rare and progressive panocular disease characterized by reduced visual acuity. Up to 90% of patients suffer from aniridia-related keratopathy (ARK), caused by a combination of factors including limbal epithelial stem cell (LSC) deficiency, impaired healing response and abnormal differentiation of the corneal epithelium. It usually begins in the first decade of life, resulting in recurrent corneal erosions, sub-epithelial fibrosis, and corneal opacification. Unfortunately, there are currently no efficient treatments available for these patients and no in vitro model for this pathology. We used CRISPR\/Cas9 technology to introduce into the PAX6 gene of LSCs a heterozygous nonsense mutation found in ARK patients. Nine clones carrying a p.E109X mutation on one allele were obtained with no off-target mutations. Compared with the parental LSCs, heterozygous mutant LSCs displayed reduced expression of PAX6 and marked slow-down of cell proliferation, migration and detachment. Moreover, addition to the culture medium of recombinant PAX6 protein fused to a cell penetrating peptide was able to activate the endogenous PAX6 gene and to rescue phenotypic defects of mutant LSCs, suggesting that administration of such recombinant PAX6 protein could be a promising therapeutic approach for aniridia-related keratopathy. More generally, our results demonstrate that introduction of disease mutations into LSCs by CRISPR\/Cas9 genome editing allows the creation of relevant cellular models of ocular disease that should greatly facilitate screening of novel therapeutic approaches.<\/jats:p>","DOI":"10.1002\/stem.2858","type":"journal-article","created":{"date-parts":[[2018,5,29]],"date-time":"2018-05-29T14:27:39Z","timestamp":1527604059000},"page":"1421-1429","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":55,"title":["Modeling of Aniridia-Related Keratopathy by CRISPR\/Cas9 Genome Editing of Human Limbal Epithelial Cells and Rescue by Recombinant PAX6 Protein"],"prefix":"10.1093","volume":"36","author":[{"given":"Lauriane N.","family":"Roux","sequence":"first","affiliation":[{"name":"INSERM U976, H\u00f4pital Saint-Louis, Paris, France"},{"name":"Universit\u00e9 Paris-Diderot, Sorbonne Paris Cit\u00e9, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabelle","family":"Petit","sequence":"additional","affiliation":[{"name":"INSERM U976, H\u00f4pital Saint-Louis, Paris, France"},{"name":"Universit\u00e9 Paris-Diderot, Sorbonne Paris Cit\u00e9, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romain","family":"Domart","sequence":"additional","affiliation":[{"name":"INSERM U1154, CNRS UMR 7196, Museum National d'Histoire Naturelle, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Paul","family":"Concordet","sequence":"additional","affiliation":[{"name":"INSERM U1154, CNRS UMR 7196, Museum National d'Histoire Naturelle, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jieqiong","family":"Qu","sequence":"additional","affiliation":[{"name":"Department of Human Developmental biology, Radboud University, Nijmegen, The Netherlands"},{"name":"Faculty of Sciences, Department of Human Genetics, Radboud University Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huiqing","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Human Developmental biology, Radboud University, Nijmegen, The Netherlands"},{"name":"Faculty of Sciences, Department of Human Genetics, Radboud University Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Joliot","sequence":"additional","affiliation":[{"name":"Coll\u00e8ge de France CNRS\/UMR 7241 - INSERM U1050, Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivier","family":"Ferrigno","sequence":"additional","affiliation":[{"name":"UMRS 938, Centre de Recherche Saint-Antoine, Paris, 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