UGT1A1 variants c.864+5G>T and c.996+2_996+5del of a Crigler-Najjar patient induce aberrant splicing in minigene assays
| العنوان: | UGT1A1 variants c.864+5G>T and c.996+2_996+5del of a Crigler-Najjar patient induce aberrant splicing in minigene assays |
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| المؤلفون: | Dmitrijs Rots, Madara Kreile, Alberto Valenzuela-Palomo, Eladio Velasco, Linda Gailite, Lara Sanoguera-Miralles |
| المساهمون: | Instituto de Salud Carlos III, Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), Junta de Castilla y León, European Commission, Asociación Española Contra el Cáncer |
| المصدر: | Digital.CSIC. Repositorio Institucional del CSIC instname Frontiers in Genetics, Vol 11 (2020) Frontiers in Genetics |
| بيانات النشر: | Frontiers Media, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Gene isoform, lcsh:QH426-470, aberrant splicing, 03 medical and health sciences, Exon, 0302 clinical medicine, Genetics, minigene, Crigler-Najjar, Aberrant splicing, Genetics (clinical), Chemistry, Intron, RNA, Brief Research Report, splice site, Molecular biology, unconjugated hyperbilirubinemia, lcsh:Genetics, 030104 developmental biology, 030220 oncology & carcinogenesis, RNA splicing, Molecular Medicine, Trans-acting, UGT1A1, Minigene |
| الوصف: | A large fraction of DNA variants impairs pre-mRNA splicing in human hereditary disorders. Crigler-Najjar syndrome (CNS) is characterized by a severe unconjugated hyperbilirubinemia caused by variants in the UGT1A1 gene. We previously reported one CNS-type II patient with two splice-site variants in trans (c.864+5G>T and c.996+2_996+5del). According to MaxEntScan, both disrupt their corresponding donor sites (c.864+5G>T: 6.99 → 2.28; c.996+2_996+5del: 5.96 → −11.02), so they were selected for subsequent functional tests. Given the unavailability of patient RNA, we constructed an UGT1A1 splicing-reporter minigene with exons 1–4 to characterize the underlying splicing anomaly. The variant c.996+2_996+5del generated two aberrant transcripts, Δ(E2) (exon 2 skipping/64%) and ▼(E2q135) (intron retention of 135-nt/36%), which lead to the loss of 18 conserved amino-acids and the gain of 45 new ones of a critical functional domain, respectively. The c.864+5G>T variant mainly produced the aberrant transcript Δ(E1q141) (141-nt deletion/70.4%) and the full-length isoform (29.6%). Δ(E1q141) would provoke the loss of 47 amino-acids of the N-terminal domain that encodes for substrate specificity. Thus, the three anomalous transcripts are likely to inactivate UGT1A1. Moreover, this patient is also homozygous for the promoter variant A(TA)7TAA that decreases UGT1A1 expression by 70%, so the full-length transcript produced by c.864+5G>T would be even more reduced ( The Scientific Laboratory of Molecular Genetics was funded by internal grants from the Riga Stradins University. EV was supported by grants from the Spanish Ministry of Science, Innovation and Universities, Plan Nacional de I + D + I 2013-2016, ISCIII (PI17/00227) co-funded by FEDER from Regional Development European Funds (European Union), and grant CSI242P18 (actuación cofinanciada P.O. FEDER 2014-2020 de Castilla y León) from the Consejería de Educación, Junta de Castilla y León. AV-P was supported by a predoctoral fellowship from the Consejería de Educación, Junta de Castilla y León (2018–2022). LS-M was supported by a predoctoral fellowship from the Spanish Association Against Cancer (AECC), Junta Provincial de Valladolid (2019–2023). |
| اللغة: | English |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::66b0c63fe40cf499be58f3171c040901 http://hdl.handle.net/10261/223205 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....66b0c63fe40cf499be58f3171c040901 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |