{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T10:01:40Z","timestamp":1753869700346,"version":"3.41.2"},"reference-count":53,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,5,9]],"date-time":"2024-05-09T00:00:00Z","timestamp":1715212800000},"content-version":"vor","delay-in-days":8,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005756","name":"Shahrekord University of Medical Sciences","doi-asserted-by":"publisher","award":["3834"],"award-info":[{"award-number":["3834"]}],"id":[{"id":"10.13039\/501100005756","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molec Gen &amp;amp; Gen Med"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>FCSK<\/jats:italic>\u2010congenital disorder of glycosylation (FCSK\u2010CDG) is a recently discovered rare autosomal recessive genetic disorder with defective fucosylation due to mutations in the fucokinase encoding gene, <jats:italic>FCSK<\/jats:italic>. Despite the essential role of fucokinase in the fucose salvage pathway and severe multisystem manifestations of FCSK\u2010CDG patients, it is not elucidated which cells or which types of fucosylation are affected by its deficiency.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study, CRISPR\/Cas9 was employed to construct an FCSK\u2010CDG cell model and explore the molecular mechanisms of the disease by lectin flow cytometry and real\u2010time PCR analyses.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Comparison of cellular fucosylation by lectin flow cytometry in the created CRISPR\/Cas9 <jats:italic>FCSK<\/jats:italic> knockout and the same unedited cell lines showed no significant change in the amount of cell surface fucosylated glycans, which is consistent with the only documented previous study on different cell types. It suggests a probable effect of this disease on secretory glycoproteins. Investigating O\u2010fucosylation by analysis of the <jats:italic>NOTCH3<\/jats:italic> gene expression as a potential target revealed a significant decrease in the <jats:italic>FCSK<\/jats:italic> knockout cells compared with the same unedited ones, proving the effect of fucokinase deficiency on EGF\u2010like repeats O\u2010fucosylation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study expands insight into the FCSK\u2010CDG molecular mechanism; to the best of our knowledge, it is the first research conducted to reveal a gene whose expression level alters due to this disease.<\/jats:p><\/jats:sec>","DOI":"10.1002\/mgg3.2445","type":"journal-article","created":{"date-parts":[[2024,5,9]],"date-time":"2024-05-09T12:39:16Z","timestamp":1715258356000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Novel insight into <i>FCSK<\/i>\u2010congenital disorder of glycosylation through a <scp>CRISPR<\/scp>\u2010generated cell model"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-7410-1254","authenticated-orcid":false,"given":"Maryam","family":"Fazelzadeh\u00a0Haghighi","sequence":"first","affiliation":[{"name":"Department of Molecular Medicine, School of Advanced Technologies Shahrekord University of Medical Sciences  Shahrekord Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7703-7387","authenticated-orcid":false,"given":"Hossein","family":"Jafari\u00a0Khamirani","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics Shiraz University of Medical Sciences  Shiraz Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8485-9247","authenticated-orcid":false,"given":"Jafar","family":"Fallahi","sequence":"additional","affiliation":[{"name":"Molecular Medicine Department, School of Advanced Medical Sciences and 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