{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T21:39:51Z","timestamp":1780349991326,"version":"3.54.1"},"reference-count":37,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2019,1,28]],"date-time":"2019-01-28T00:00:00Z","timestamp":1548633600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2019,3]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Classically, the study of splicing impact of variation located near the splice site is performed by both in silico and mRNA analysis. However, RNA sample was rarely available.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To characterize a panel of putative haemophilia A splicing variations.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>Twenty\u2010six <jats:italic>F8 <\/jats:italic>variations identified from a cohort of 2075 haemophilia A families were studied using both bioinformatic tools and in vitro minigene assays in HeLa and Huh7 cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>An aberrant splicing was demonstrated for 21\/26 tested sequence variations. A good correlation between in silico and in vitro analysis was obtained for variations affecting donor splice site (12\/14) and for the synonymous variations located inside an exon (6\/6). Conversely, no concordant results were observed for the six variations affecting acceptor splice sites. The variations resulted more frequently in exon skipping (n\u00a0=\u00a013) than in activation of nearby cryptic splice sites (n\u00a0=\u00a05), in use of a de novo splice site (n\u00a0=\u00a02) or in insertion of large intronic sequences (n\u00a0=\u00a01). This study allowed to reclassify 5 synonymous substitutions c.1167A&gt;G (p.Gln389Gln), c.1569G&gt;T (p.Leu523Leu), c.1752G&gt;A (p.Gln584Gln), c.5586G&gt;A (p.Leu1862Leu) and c.6066C&gt;T (p.Gly2022Gly) as splicing variations. The pathological significance of five variations remained unclear (c.222G&gt;A [p.Thr74Thr], c.237C&gt;T [p.Asn79Asn], c.240C&gt;T [p.Ile80Ile], c.2113+5_2113+8del and c.2113+5G&gt;A).<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The minigene assay herein gave additional evidences for the clinical significance of 21\/26 <jats:italic>F8 <\/jats:italic>putative splice site mutations. Such investigation should be performed for each <jats:italic>F8<\/jats:italic> putative splice site variation for which no mRNA sample is available, notably to greatly improve the genetic counselling given to female carriers.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hae.13687","type":"journal-article","created":{"date-parts":[[2019,1,29]],"date-time":"2019-01-29T04:00:34Z","timestamp":1548734434000},"page":"306-315","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Splicing analysis of 26 <i>F8<\/i> nucleotide variations using a minigene assay"],"prefix":"10.1111","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7408-6474","authenticated-orcid":false,"given":"Yohann","family":"Jourdy","sequence":"first","affiliation":[{"name":"Service d\u2019H\u00e9matologie Biologique, Centre de Biologie et Pathologie Est Hospices Civils de Lyon Lyon France"},{"name":"EA 4609 H\u00e9mostase et Cancer, Universit\u00e9 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