{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T19:40:44Z","timestamp":1790278844739,"version":"4.1.0"},"reference-count":86,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T00:00:00Z","timestamp":1676937600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Slovenian Research Agency","award":["P3-0124"],"award-info":[{"award-number":["P3-0124"]}]},{"name":"Slovenian Research Agency","award":["J3-8207"],"award-info":[{"award-number":["J3-8207"]}]},{"name":"Slovenian Research Agency","award":["MR51882"],"award-info":[{"award-number":["MR51882"]}]},{"name":"Slovenian Research Agency","award":["OP20.05187 RI-SI-EATRIS"],"award-info":[{"award-number":["OP20.05187 RI-SI-EATRIS"]}]},{"name":"the Ministry of Education, Science, and Sport (MIZ\u0160) and the European Regional Development Fund","award":["P3-0124"],"award-info":[{"award-number":["P3-0124"]}]},{"name":"the Ministry of Education, Science, and Sport (MIZ\u0160) and the European Regional Development Fund","award":["J3-8207"],"award-info":[{"award-number":["J3-8207"]}]},{"name":"the Ministry of Education, Science, and Sport (MIZ\u0160) and the European Regional Development Fund","award":["MR51882"],"award-info":[{"award-number":["MR51882"]}]},{"name":"the Ministry of Education, Science, and Sport (MIZ\u0160) and the European Regional Development Fund","award":["OP20.05187 RI-SI-EATRIS"],"award-info":[{"award-number":["OP20.05187 RI-SI-EATRIS"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Although the aetiology of non-syndromic orofacial clefts (nsOFCs) is usually multifactorial, syndromic OFCs (syOFCs) are often caused by single mutations in known genes. Some syndromes, e.g., Van der Woude syndrome (VWS1; VWS2) and X-linked cleft palate with or without ankyloglossia (CPX), show only minor clinical signs in addition to OFC and are sometimes difficult to differentiate from nsOFCs. We recruited 34 Slovenian multi-case families with apparent nsOFCs (isolated OFCs or OFCs with minor additional facial signs). First, we examined IRF6, GRHL3, and TBX22 by Sanger or whole exome sequencing to identify VWS and CPX families. Next, we examined 72 additional nsOFC genes in the remaining families. Variant validation and co-segregation analysis were performed for each identified variant using Sanger sequencing, real-time quantitative PCR and microarray-based comparative genomic hybridization. We identified six disease-causing variants (three novel) in IRF6, GRHL3, and TBX22 in 21% of families with apparent nsOFCs, suggesting that our sequencing approach is useful for distinguishing syOFCs from nsOFCs. The novel variants, a frameshift variant in exon 7 of IRF6, a splice-altering variant in GRHL3, and a deletion of the coding exons of TBX22, indicate VWS1, VWS2, and CPX, respectively. We also identified five rare variants in nsOFC genes in families without VWS or CPX, but they could not be conclusively linked to nsOFC.<\/jats:p>","DOI":"10.3390\/ijms24054262","type":"journal-article","created":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T02:30:35Z","timestamp":1676946635000},"page":"4262","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["A Comprehensive Genetic Analysis of Slovenian Families with Multiple Cases of Orofacial Clefts Reveals Novel Variants in the Genes IRF6, GRHL3, and TBX22"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5904-8939","authenticated-orcid":false,"given":"Lara","family":"Slavec","sequence":"first","affiliation":[{"name":"Research Unit, Division of Gynaecology and Obstetrics, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9088-2382","authenticated-orcid":false,"given":"Ksenija","family":"Ger\u0161ak","sequence":"additional","affiliation":[{"name":"Research Unit, Division of Gynaecology and Obstetrics, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Department of Gynaecology and Obstetrics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreja","family":"Eberlinc","sequence":"additional","affiliation":[{"name":"Department of Maxillofacial and Oral Surgery, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5330-3983","authenticated-orcid":false,"given":"Tinka","family":"Hovnik","sequence":"additional","affiliation":[{"name":"Clinical Institute for Special Laboratory Diagnostics, University Children\u2019s Hospital, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Lovre\u010di\u0107","sequence":"additional","affiliation":[{"name":"Department of Gynaecology and Obstetrics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Clinical Institute of Genomic Medicine, University Medical Centre Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irena","family":"Mlinari\u010d-Ra\u0161\u010dan","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nata\u0161a","family":"Karas Ku\u017eeli\u010dki","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,21]]},"reference":[{"key":"ref_1","first-page":"1","article-title":"Epidemiological Studies on the Frequency of Clefts in Europe and World-Wide","volume":"34","author":"Gundlach","year":"2006","journal-title":"J. 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