{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T11:16:53Z","timestamp":1772795813164,"version":"3.50.1"},"reference-count":48,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T00:00:00Z","timestamp":1772755200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007086","name":"Sel\u00e7uk University","doi-asserted-by":"publisher","award":["19102037"],"award-info":[{"award-number":["19102037"]}],"id":[{"id":"10.13039\/501100007086","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Over 70 mutations in the transforming growth factor beta-induced (TGFBI) gene are associated with corneal dystrophies that impair vision. The R555W hotspot mutation is a major cause of granular corneal dystrophy type 1 (GCD1). Here, we evaluated the technical feasibility of CRISPR\/Cas9-mediated editing of the R555W mutation in peripheral blood mononuclear cells (PBMCs) obtained from a patient with GCD1. Three single guide RNAs (sgRNA1\u20133) and matched single-stranded oligodeoxynucleotide donors (ssODN1\u20133) were designed and co-transfected into PBMCs. Transfected cells were enriched by flow cytometric sorting, with GFP-positive cells representing approximately 2\u20134% of the total electroporated population. Editing outcomes were initially screened using high-resolution melting (HRM) analysis, and the sgRNA3\u2013ssODN3 combination identified as the most promising candidate was subsequently validated by next-generation sequencing (NGS). Sequencing revealed a homology-directed repair efficiency of 98.2% among GFP-positive sorted cells, demonstrating efficient and precise genome editing within the enriched population. Because PBMCs are not disease-relevant corneal epithelial cells and only genomic endpoints were assessed, the clinical applicability of this study is limited and the work should be considered a technical proof-of-concept. This framework supports optimization of CRISPR-based strategies prior to studies in biologically relevant corneal models.<\/jats:p>","DOI":"10.3390\/ijms27052418","type":"journal-article","created":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T09:22:14Z","timestamp":1772788934000},"page":"2418","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Precise CRISPR-Mediated Editing of the TGFBI R555W Mutation in Patient-Derived Peripheral Blood Mononuclear Cells"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4362-3852","authenticated-orcid":false,"given":"Burak","family":"Dagdelen","sequence":"first","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Sel\u00e7uk University, 42130 Konya, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6600-6603","authenticated-orcid":false,"given":"Hilal","family":"Arikoglu","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Sel\u00e7uk University, 42130 Konya, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dudu","family":"Erkoc-Kaya","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, Sel\u00e7uk University, 42130 Konya, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9847-3521","authenticated-orcid":false,"given":"Banu","family":"Bozkurt","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Faculty of Medicine, Sel\u00e7uk University, 42130 Konya, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e177","DOI":"10.1016\/j.jfo.2017.05.003","article-title":"Corneal dystrophies","volume":"40","author":"Bourges","year":"2017","journal-title":"J. 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Release"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2418\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T09:23:48Z","timestamp":1772789028000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/5\/2418"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,6]]},"references-count":48,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["ijms27052418"],"URL":"https:\/\/doi.org\/10.3390\/ijms27052418","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,6]]}}}