BUSCADOR RECOLECTA

Digital.CSIC. Repositorio Institucional del CSIC

oai:digital.csic.es:10261/278888

[Method] Splicing functional assays of PALB2 variants by hybrid minigenes. Protocol Workflow: 1) Minigene construction. 2) Site-directed mutagenesis. 3) Cell Transfection. 4) RNA Isolation. 5) RT-PCR. 6) Transcript characterization (Sanger sequencing and fluorescent fragment electrophoresis)., This dataset corresponds to a comprehensive splicing analysis of splice-site variants of exons 1 to 3 of the breast cancer susceptibility gene PALB2. These variants were reported at the ClinVar database. Loss-of-function variants at the PALB2 gene are known to confer risk to breast and ovarian cancers. A total of 31 PALB2 variants at the intron/exon boundaries were analyzed with MaxEntScan. Sixteen variants were selected and genetically engineered into a PALB2 splicing reporter minigene. We found that 12 variants disrupted splicing and six of them could be classified as likely pathogenic. Hence, they are clinically actionable findings so variant-carriers may benefit from tailored prevention protocols and therapies., This work was supported by grants from the Spanish Ministry of Science and Innovation, Plan Nacional de I+D+I 2013-2016, ISCIII (PI20/00225) co-funded by FEDER from Regional Development European Funds (European Union) and from the Consejería de Educación, Junta de Castilla y León, ref. CSI242P18 (actuación cofinanciada P.O. FEDER 2014-2020 de Castilla y León)., Peer reviewed