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Onc."],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Pathogenic germline variants in\n                    <jats:italic>SDHB<\/jats:italic>\n                    predispose to pheochromocytoma and paraganglioma, but limited functional evidence challenges clinical interpretation. To investigate splice-associated\n                    <jats:italic>SDHB<\/jats:italic>\n                    variants, we developed a minigene spanning exons 2-5 and assessed derived\n                    <jats:italic>SDHB<\/jats:italic>\n                    transcripts in HEK293T cells using targeted RNA sequencing. We evaluated 48 variants prioritized by SpliceAI (\u0394\u22650.42), two negative controls and endogenous\n                    <jats:italic>SDHB<\/jats:italic>\n                    , and compared findings with tumor data (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u20092). Nineteen variants (38%) showed \u226590% wildtype splicing, whereas 17 (34%) exhibited \u226590% aberrant splicing. Across all variants, 73 aberrant transcripts were observed (average of 2.3 per variant, 22 unique transcripts). Using a customized decision framework, RNA-based evidence strengths were assigned to 64 aberrant transcripts (88%). Among 26 classified variants, 10 received PVS1_Strong (RNA) (38%), including eight canonical splice-site variants, one missense variant and one stop-gain variant; two received PVS1_Moderate (RNA) and 14 received BP7_Strong (RNA). Integration of minigene RNA data changed ACMG scores by a mean of 2.7 points and led to reclassification of 13 variants (50%), including 12 downgrades from VUS to likely benign and one downgrade from likely pathogenic to VUS. These findings demonstrate that targeted sequencing of minigene-derived transcripts provides a scalable approach to evaluate splice-associated\n                    <jats:italic>SDHB<\/jats:italic>\n                    variants and improve variant classification.\n                  <\/jats:p>","DOI":"10.1038\/s41698-026-01685-7","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T18:24:46Z","timestamp":1788891886000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Minigene-based characterization and classification of splice-associated variants in succinate dehydrogenase B"],"prefix":"10.1038","volume":"10","author":[{"given":"Anni","family":"K\u00f6hler","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra A.","family":"Baumann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natasha","family":"Lewis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anja","family":"Richter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan","family":"Richter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doreen","family":"William","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9s","family":"Cruz-Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanno","family":"Glimm","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Fr\u00f6hling","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Le Duc","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evelin","family":"Schr\u00f6ck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arne","family":"Jahn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"1685_CR1","doi-asserted-by":"publisher","first-page":"8812","DOI":"10.1200\/JCO.2005.03.1484","volume":"23","author":"L Amar","year":"2005","unstructured":"Amar, L. et al. 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