Long-read assessment of gene transcripts for developing variant classification guidelines

Analysis of ‘naturally occurring’ mRNA splice isoforms for breast cancer susceptibility genes aids in the interpretation of in vitro splicing assays for sequence variant classification. We have been applying long-read nanopore sequencing, short-read RNA-seq (whole transcriptome and targeted), and PCR-based technologies to catalogue alternative mRNA transcripts from breast and ovarian cancer susceptibility genes. We are using these data to evaluate potential pathogenicity of gene variants located at canonical splice sites and refine the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG/AMP) variant classification guidelines.