A Yeast-Based High-Throughput Screening Platform for the Discovery of Novel pre-mRNA Splicing Modulators

Area of Science:

• Molecular Biology
• Genetics
• Drug Discovery

Background:

• Pre-messenger RNA (mRNA) splicing is crucial for eukaryotic gene expression.
• Dysregulation of splicing is implicated in numerous diseases.
• Few small molecules exist to modulate splicing or inhibit splicing machinery.

Purpose of the Study:

• To develop and validate a novel high-throughput screening (HTS) platform for identifying splicing-modulating compounds.
• To discover small molecules that can inhibit or alter pre-mRNA splicing.
• To investigate the therapeutic potential of identified compounds in cancer models.

Main Methods:

• A two-tiered screening approach using sensitized yeast strains and fluorescent protein production.
• Primary screen: measuring growth inhibition in yeast.
• Secondary screen: assessing splicing inhibition via fluorescent reporter assay.

Main Results:

• Identified four small molecules that induce pre-mRNA accumulation in yeast.
• One compound demonstrated increased sensitivity in cancer cells with SRSF2^P95H mutation compared to wild-type.
• Transcriptome analysis revealed widespread gene expression changes in sensitive cells.

Conclusions:

• A yeast-based HTS platform is effective for discovering splicing modulators.
• Identified compounds represent potential therapeutic agents for splicing-related diseases.
• The study highlights the utility of yeast models for drug discovery targeting splicing.