biostudies-literatureValenzuela-Palomo AAsociación Española Contra el CáncerInstituto de Salud Carlos IIIJunta de Castilla y LeónComunidad de MadridUniversity of Valladolid4541https://www.ebi.ac.uk/biostudies/studies/S-EPMC9496955biostudies-literatureUnknown14(18)<i>PALB2</i> loss-of-function variants are associated with significant increased risk of breast cancer as well as other types of tumors. Likewise, splicing disruptions are a common mechanism of disease susceptibility. Indeed, we previously showed, by minigene assays, that 35 out of 42 <i>PALB2</i> variants impaired splicing. Taking advantage of one of these constructs (mgPALB2_ex1-3), we proceeded to analyze other variants at exons 1 to 3 reported at the ClinVar database. Thirty-one variants were bioinformatically analyzed with MaxEntScan and SpliceAI. Then, 16 variants were selected for subsequent RNA assays. We identified a total of 12 spliceogenic variants, 11 of which did not produce any trace of the expected minigene full-length transcript. Interestingly, variant c.49-1G &gt; A mimickCancersSplicing Analysis of 16 <i>PALB2</i> ClinVar Variants by Minigene Assays: Identification of Six Likely Pathogenic Variants.PMC9496955Predoctoral fellowship IL-BAE-S: Operational Program for Youth Employment and the Youth Employment InitiativePI20/00110Postdoctoral researcher EB-M (POSTDOC-UVA05, 2022-2025)Predoctoral fellowship LS-MPI20/00225CSI242P18Velasco-Sampedro EASanoguera-Miralles LEsteban-Sanchez APerez-Segura PLlinares-Burguet IGomez-Barrero SGarcia-Alvarez AValenzuela-Palomo ABueno-Martinez Ede la Hoya MfalseSplicing Analysis of 16 <i>PALB2</i> ClinVar Variants by Minigene Assays: Identification of Six Likely Pathogenic Variants.<i>PALB2</i> loss-of-function variants are associated with significant increased risk of breast cancer as well as other types of tumors. Likewise, splicing disruptions are a common mechanism of disease susceptibility. Indeed, we previously showed, by minigene assays, that 35 out of 42 <i>PALB2</i> variants impaired splicing. Taking advantage of one of these constructs (mgPALB2_ex1-3), we proceeded to analyze other variants at exons 1 to 3 reported at the ClinVar database. Thirty-one variants were bioinformatically analyzed with MaxEntScan and SpliceAI. Then, 16 variants were selected for subsequent RNA assays. We identified a total of 12 spliceogenic variants, 11 of which did not produce any trace of the expected minigene full-length transcript. Interestingly, variant c.49-1G &gt; A mimick2022-01-01T00:00:00Z2022 Sep2025-04-22T00:36:54.713Z2025-04-05T19:38:42.087ZS-EPMC94969553613969910.3390/cancers14184541