ParSE-seq: a calibrated multiplexed assay to facilitate the clinical classification of putative splice-altering variants.

ABSTRACT: Interpreting the clinical significance of putative splice-altering variants outside canonical splice sites remains difficult without time-intensive experimental studies. To address this, we introduce Parallel Splice Effect Sequencing (ParSE-seq), a multiplexed assay to quantify variant effects on RNA splicing. We first apply this technique to study hundreds of variants in the arrhythmia-associated gene SCN5A. Variants are studied in 'minigene' plasmids with molecular barcodes to allow pooled variant effect quantification. We perform experiments in two cell types, including disease-relevant induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). The assay strongly separates known control variants from ClinVar, enabling quantitative calibration of the ParSE-seq assay. Using these ev

SUBMITTER: O'Neill MJ 

PROVIDER: S-EPMC11437130 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature