Deleterious mutations in BRCA1 and BRCA2 increased up to 20-fold the risk of developing breast cancer. Previously published data of our group showed that a third part of pathogenic mutations affected pre-mRNA processing so that splicing could be one of the most relevant etiopathogenic mechanism in hereditary breast/ovarian cancer. We constructed a hybrid minigene with BRCA1 exons 15 to 19 in the new splicing vector pSAD (Spanish patent: P201231427). Bioinformatics studies of DNA variants in exons 16 and 17 of BRCA1 were performed with NNSplice and Human Splicing Finder, to select them according to the following criteria: disruption of canonical splice sites, broken branch points or creation of de novo silencers. Functional assays performed either in lymphocyte RNA from patients or in hybrid minigenes in MCF7/HeLa cells. We investigated the impact on splicing of pre-selected variants listed in the international databases and to correlate these results with those obtained in lymphocyte RNA of patients from Castilla y Leon (Spain). All the RT-PCR products were sequenced to characterize all the splicing outcomes. Splicing functional assays allow a better molecular characterization of DNA variants of unknown clinical significance. The ultimate goal of this research is to improve the quality of life of patients and their families that will benefit from new preventive measures, surveillance and prophylaxis.

Resumen del póster presentado a la European Human Genetics Conference celebrada en Paris (Francia) del 9 al 11 de junio de 2013.

Peer Reviewed