• Folder: 1.Sequences - Sub-folder cDNA. Transcript Sequencing. Sub-folders: - mgATM_11-17: 8 *.ab1 files of transcripts generated by variants introduced into minigene mgATM_11-17 - mgATM_41-44: 12 *.ab1 files of transcripts generated by variants introduced into minigene mgATM_41-44 - mgBRCA1_13-19: 17 *.ab1 files of transcripts generated by variants introduced into minigene mgBRCA1_13-19 - mgPALB2_1-3: 7 *.ab1 files of transcripts generated by variants introduced into minigene mgPALB2_1-3 - mgPALB2_5-12: 18 *.ab1 files of transcripts generated by variants introduced into minigene mgPALB2_5-12 - Sub-folder: Minigenes. Sequence files of wild type and mutant constructs. 51 *.ab1 files. - mgATM_11-17: 7 *.ab1 files. - mgATM_41-44: 8 *.ab1 files. - mgBRCA1_13-19: 16 *.ab1 files. - mgPALB2_1-3: 5 *.ab1 files. - mgPALB2_5-12: 15 *.ab1 files. • Folder: 2.Fragment_Analysis. Fluorescent Fragment Analysis of plicing assays. Sub-folders: - mgATM_11-17: 12 *.fsa files of fluorescent fragment analysis of RT-PCRs of variants. - mgATM_41-44: 27 *.fsa files of fluorescent fragment analysis of RT-PCRs of variants. - mgBRCA1_13-19: 33 *.fsa files of fluorescent fragment analysis of RT-PCRs of variants. - mgPALB2_1-3: 12 *.fsa files of fluorescent fragment analysis of RT-PCRs of variants. - mgPALB2_5-12: 28 *.fsa files of fluorescent fragment analysis of RT-PCRs of variants.

This dataset contains fragment analysis and sequencing files of the impact on splicing of +2T variants of the breast cancer susceptibility genes. Splicing dysregulation is a well-established mechanism of pathogenicity for variants in disease susceptibility genes. Variants at the critical intronic +1,2 GT nucleotides of the 5’ splice-site (5’ss) are considered strong signals of pathogenicity. However, some +2T variants create functional non-canonical 5’ss that generate wild-type transcripts, hampering accurate variant interpretation and genetic counselling. We investigated the functional impact of 29 +2T>N variants of the breast cancer susceptibility genes ATM, BRCA1 and PALB2 using minigene splicing assays. Six variants generated wild-type transcripts (4%-81% of the overall expression). As expected, high expression of wild-type transcripts significantly affected variant interpretation.

EAV-S lab is supported by grants from the Spanish Ministry of Science and Innovation, Instituto de Salud Carlos III (PI23/00047) co-funded by FEDER from Regional Development European Funds (European Union).

[Description of methods used for collection/generation of data] - Sanger sequencing - Fluorescent Fragment Analysis

Peer reviewed