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dc.contributor.authorSanoguera-Miralles, Lara
dc.contributor.authorValenzuela-Palomo, Alberto
dc.contributor.authorBueno-Martínez, Elena
dc.contributor.authorLlovet, Patricia
dc.contributor.authorDíez-Gómez, Beatriz
dc.contributor.authorCaloca, María José
dc.contributor.authorPérez-Segura, Pedro
dc.contributor.authorFraile-Bethencourt, Eugenia
dc.contributor.authorColmena, Marta
dc.contributor.authorCarvalho, Sara
dc.contributor.authorAllen, Jamie
dc.contributor.authorEaston, Douglas F.
dc.contributor.authorDevilee, Peter
dc.contributor.authorVreeswijk, Maaike P. G.
dc.contributor.authorde la Hoya, Miguel
dc.contributor.authorVelasco, Eladio A.
dc.date.accessioned2020-12-22T18:56:26Z
dc.date.available2020-12-22T18:56:26Z
dc.date.issued2020-12-15
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/315427
dc.description.abstractHereditary breast and/or ovarian cancer is a highly heterogeneous disease with more than 10 known disease-associated genes. In the framework of the BRIDGES project (Breast Cancer Risk after Diagnostic Gene Sequencing), the RAD51C gene has been sequenced in 60,466 breast cancer patients and 53,461 controls. We aimed at functionally characterizing all the identified genetic variants that are predicted to disrupt the splicing process. Forty RAD51C variants of the intron-exon boundaries were bioinformatically analyzed, 20 of which were selected for splicing functional assays. To test them, a splicing reporter minigene with exons 2 to 8 was designed and constructed. This minigene generated a full-length transcript of the expected size (1062 nucleotides), sequence, and structure (Vector exon V1- RAD51C exons_2-8- Vector exon V2). The 20 candidate variants were genetically engineered into the wild type minigene and functionally assayed in MCF-7 cells. Nineteen variants (95%) impaired splicing, while 18 of them produced severe splicing anomalies. At least 35 transcripts were generated by the mutant minigenes: 16 protein-truncating, 6 in-frame, and 13 minor uncharacterized isoforms. According to ACMG/AMP-based standards, 15 variants could be classified as pathogenic or likely pathogenic variants: c.404G > A, c.405-6T > A, c.571 + 4A > G, c.571 + 5G > A, c.572-1G > T, c.705G > T, c.706-2A > C, c.706-2A > G, c.837 + 2T > C, c.905-3C > G, c.905-2A > C, c.905-2_905-1del, c.965 + 5G > A, c.1026 + 5_1026 + 7del, and c.1026 + 5G > T.
dc.languageen
dc.publisherMDPI
dc.subjectbreast cancer
dc.subjectovarian cancer
dc.subjectsusceptibility genes
dc.subjectRAD51C
dc.subjectgenetic variants
dc.subjectsplicing
dc.subjectaberrant splicing
dc.subjectVUS
dc.subjectfunctional assay
dc.subjectminigene
dc.subjectclinical interpretation
dc.titleComprehensive Functional Characterization and Clinical Interpretation of 20 Splice-Site Variants of the RAD51C Gene
dc.typeArticle
dc.date.updated2020-12-22T18:56:25Z
prism.issueIdentifier12
prism.publicationNameCancers
prism.volume12
dc.identifier.doi10.17863/CAM.62534
dcterms.dateAccepted2020-12-09
rioxxterms.versionofrecord10.3390/cancers12123771
rioxxterms.versionVoR
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0/
dc.contributor.orcidSanoguera-Miralles, Lara [0000-0002-5548-7114]
dc.contributor.orcidBueno-Martínez, Elena [0000-0003-4888-2885]
dc.contributor.orcidLlovet, Patricia [0000-0003-0190-2105]
dc.contributor.orcidFraile-Bethencourt, Eugenia [0000-0001-8828-5992]
dc.contributor.orcidDevilee, Peter [0000-0002-8023-2009]
dc.contributor.orcidVreeswijk, Maaike P. G. [0000-0003-4068-9271]
dc.contributor.orcidde la Hoya, Miguel [0000-0002-8113-1410]
dc.contributor.orcidVelasco, Eladio A. [0000-0002-9682-5589]
dc.identifier.eissn2072-6694


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