The SeqSplice multiplexed minigene splicing assay for characterization and quantitation of variant-induced <i>BRCA1</i> and <i>BRCA2</i> splice isoforms.

ABSTRACT: BRCA1 and BRCA2 germline variant classification is vital for clinical management of families with hereditary breast and ovarian cancer. However, clinical classification of rare variants outside of the splice donor/acceptor ±1,2-dinucleotides remains challenging, particularly for variants that induce new or cryptic splice site usage. Here, we present SeqSplice, a high-throughput RNA splicing methodology utilizing barcoded minigene constructs together with a bespoke bioinformatics pipeline for identifying and quantifying the impacts for splice-altering variants. SeqSplice exhibits excellent reproducibility across cDNA input and PCR cycle differences and is able to identify and quantitate transcripts that differed by a single base. Of the 193 BRCA1 and 72 BRCA2<

SUBMITTER: Canson DM 

PROVIDER: S-EPMC12401047 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature