{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T10:11:11Z","timestamp":1790244671916,"version":"4.1.0"},"reference-count":38,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":266,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100009287","name":"NSW Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009287","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Human Mutation"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:p>\n                    Variants affecting minor (U12\u2010type) intron splice sites are underreported, and their consequences are poorly understood, which makes predicting their pathogenicity challenging. We investigated the impact on splicing of an\n                    <jats:italic>SCN5A<\/jats:italic>\n                    minor intron donor variant in an individual with cardiac arrhythmias using patient\u2010specific induced pluripotent stem cell cardiomyocytes (iPSC\u2010CMs). Genetic testing in the proband identified an\n                    <jats:italic>SCN5A<\/jats:italic>\n                    c.392+3A&gt;G variant of uncertain significance in a minor intron donor splice site, which segregated in affected family members with cardiac phenotypes, including Brugada syndrome. Transcriptome sequencing of proband\u2010specific iPSC\u2010CMs, treated with and without inhibition of nonsense\u2010mediated decay, revealed aberrant\n                    <jats:italic>SCN5A<\/jats:italic>\n                    splicing. Four variant\u2010associated isoforms were absent from control iPSC\u2010CMs and control myectomy samples, and one skipped Exon 3 with a 96\u2009bp extension of Exon 4. All alternatively spliced\n                    <jats:italic>SCN5A<\/jats:italic>\n                    transcripts were T\/A cloned and confirmed by Sanger sequencing. These alternatively spliced transcripts used novel combinations of cryptic minor and major splice sites and introduced premature termination codons or an in\u2010frame deletion, supporting reclassification of the variant as likely pathogenic. A genome\u2010wide analysis of SpliceAI scores across minor intron splice sites retrieved from the Minor Intron Database revealed clear patterns of variants with high predicted splicing impact. This analysis also highlighted distinct splice site sequence motifs between minor and major splice sites that are not currently considered in ACMG\/AMP variant interpretation guidelines. We further demonstrate that some cryptic donor\u2013acceptor motif pairings are likely incompatible with splicing, which may inform future improvements in in silico splice site prediction tools. Integrating functional RNA sequencing with SpliceAI\u2010based predictions at minor introns offers a pathway towards refining variant interpretation frameworks in clinical genomics.\n                  <\/jats:p>","DOI":"10.1155\/humu\/4537601","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T09:48:19Z","timestamp":1790243299000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<i>SCN5A<\/i>\n                    U12\u2010Type Minor Intron Variant Causes Cryptic Splicing and Cardiac Arrhythmia Disease"],"prefix":"10.1155","volume":"2026","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0264-3395","authenticated-orcid":false,"given":"Emma 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