HCS-Splice: A High-Content Screening Method to Advance the Discovery of RNA Splicing-Modulating Therapeutics

Nucleic acid therapeutics have demonstrated an impressive acceleration in recent years. They work through multiple mechanisms of action, including the downregulation gene expression and modulation RNA splicing. While several drugs based on former mechanism been approved, few target latter, despite promise splicing modulation. To improve our ability to discover novel splicing-modulating therapies, we developed HCS-Splice, a robust cell-based High-Content Screening (HCS) assay. By implementing use two-colour (GFP/RFP) fluorescent reporter plasmid, versatile, effective, rapid, high-throughput strategy for identification potent molecules. The HCS-Splice can also be used functionally confirm mutations human genetic disorders or screen drug candidates. As proof-of-concept, introduced dementia-related splice-switching mutation Microtubule-Associated Protein Tau (MAPT) exon 10 reporter. We applied wild-type mutant reporters measured functional change inclusion. demonstrate applicability method discovery, was evaluate efficacy 10-targeting siRNA, which able restore correct alternative balance.