Background
Every human cell must accurately read and interpret its genetic code. To do this, cells copy DNA into pre-messenger RNAs (pre-mRNAs), which are precisely edited by the spliceosome to produce mature, protein-coding mRNAs. As a dynamic, RNA-based enzyme, the spliceosome not only ensures accurate RNA processing and gene expression but also drives evolutionary plasticity. By allowing a single gene to encode multiple protein variants, alternative splicing serves as a major engine of proteomic complexity and cellular adaptability. Our previous cryo-electron microscopy work has captured the spliceosome in action, revealing how its RNA core and specific splicing factors recognize and position the splice sites during catalysis. When these dynamic structural rearrangements and specific protein-RNA interactions are perturbed, they frequently lead to pathogenic mis-splicing in various diseases.
Goals
Our primary goal is to dissect the molecular mechanics of the spliceosome's catalytic stage, transitioning from isolated in vitro splicing systems to the native in vivo chromatin environment. By combining reconstituted human spliceosomes, time-resolved cryo-EM, and quantitative biochemistry and proteomics, we aim to understand how specific splicing factors and ATPases regulate active site dynamics, splice site selection, and proofreading. We are particularly focused on how the dynamic engagement of exon-ligation factors fine-tunes alternative splicing in a transcript-specific manner during catalysis. To extend these findings genome-wide, we ultimately aim to purify and visualise native co-transcriptional spliceosome assemblies to delineate how chromatin context and tissue-specific signals govern the assembly and function of substrate-specialized spliceosomes. By mapping these regulatory mechanisms we hope to discover fundamental principles that govern gene expression and to uncover novel therapeutic targets for correcting mis-splicing in human disease.
