Diagnostic Utility of RNA Sequencing for Reclassification of Rare Disease Variants by Exome and Genome Sequencing

BACKGROUND
• Variants of uncertain significance (VUS) present significant challenges in interpretation and medical management for healthcare providers and patients.
• RNA sequencing (RNAseq), which has been used in hereditary cancer testing to aid in variant interpretation, has been increasingly used to provide functional evidence in genome sequencing (GS) and exome sequencing (ES).
• For variants expected to impact splicing and gene expression, RNAseq may allow for reclassification and greater diagnostic clarity.
• Here, we describe our diagnostic laboratory’s experience with leveraging clinically-validated RNAseq using EDTA blood to clarify the significance of variants obtained by GS/ES.