{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T22:15:16Z","timestamp":1792707316945,"version":"4.5.0"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,12,1]],"date-time":"2017-12-01T00:00:00Z","timestamp":1512086400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,12,1]],"date-time":"2017-12-01T00:00:00Z","timestamp":1512086400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Granular corneal dystrophy (GCD) is an autosomal dominant hereditary disease in which multiple discrete and irregularly shaped granular opacities are deposited in the corneal stroma. GCD is caused by a point mutation in the <jats:italic>transforming growth factor-\u03b2-induced<\/jats:italic> (<jats:italic>TGFBI<\/jats:italic>) gene, located on chromosome 5q31. Here, we report the first successful application of CRISPR-Cas9-mediated genome editing for the correction of a <jats:italic>TGFBI<\/jats:italic> mutation in GCD patient-derived primary corneal keratocytes via homology-directed repair (HDR). To correct genetic defects in GCD patient cells, we designed a disease-specific guide RNA (gRNA) targeting the R124H mutation of <jats:italic>TGFBI<\/jats:italic>, which causes GCD type 2 (GCD2). An R124H mutation in primary human corneal keratocytes derived from a GCD2 patient was corrected by delivering a CRISPR plasmid expressing Cas9\/gRNA and a single-stranded oligodeoxynucleotide HDR donor template <jats:italic>in vitro<\/jats:italic>. The gene correction efficiency was 20.6% in heterozygous cells and 41.3% in homozygous cells. No off-target effects were detected. These results reveal a new therapeutic strategy for GCD2; this method may also be applicable to other heredity corneal diseases.<\/jats:p>","DOI":"10.1038\/s41598-017-16308-2","type":"journal-article","created":{"date-parts":[[2017,11,27]],"date-time":"2017-11-27T10:49:11Z","timestamp":1511779751000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":46,"title":["Repair of the TGFBI gene in human corneal keratocytes derived from a granular corneal dystrophy patient via CRISPR\/Cas9-induced homology-directed repair"],"prefix":"10.1038","volume":"7","author":[{"given":"Yukako","family":"Taketani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kohdai","family":"Kitamoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshihiro","family":"Sakisaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikiko","family":"Kimakura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsuya","family":"Toyono","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoru","family":"Yamagami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiro","family":"Amano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masahiko","family":"Kuroda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tara","family":"Moore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomohiko","family":"Usui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuo","family":"Ouchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,12,1]]},"reference":[{"key":"16308_CR1","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1097\/ICO.0000000000000307","volume":"34","author":"JS Weiss","year":"2015","unstructured":"Weiss, J. 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Clinically atypical granular corneal dystrophy with pathologic features of lattice-like amyloid deposits. A study of three families. Ophthalmology \n                           95, 46\u201351 (1988).","journal-title":"Ophthalmology"},{"key":"16308_CR7","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1001\/archopht.1982.01030030792013","volume":"100","author":"PJ Dubord","year":"1982","unstructured":"Dubord, P. J. & Krachmer, J. H. Diagnosis of early lattice corneal dystrophy. Arch. Ophthalmol. \n                           100, 788\u2013790 (1982).","journal-title":"Arch. Ophthalmol."},{"key":"16308_CR8","doi-asserted-by":"publisher","first-page":"1490","DOI":"10.1016\/S0161-6420(99)90441-4","volume":"106","author":"R Dinh","year":"1999","unstructured":"Dinh, R., Rapuano, C. J., Cohen, E. J. & Laibson, P. R. Recurrence of corneal dystrophy after excimer laser phototherapeutic keratectomy. 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PLoS One \n                           10, e0116838 (2015).","journal-title":"PLoS One"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-16308-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-16308-2","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-16308-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T14:41:11Z","timestamp":1671806471000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-16308-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,12,1]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["16308"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-16308-2","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,12,1]]},"assertion":[{"value":"28 February 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 November 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 December 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Dr. Tomohiko Usui has had financial support from Santen Pharmaceutical Co., Ltd., Senju Pharmaceutical Co., Ltd., and Alcon, Inc. The other authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"16713"}}