Decoding subcellular spatial biology with high precision using RNA photocatalysts

Introduction

RNA is a fundamental component of life. Complex, dynamic, and spatial networks of molecular interactions between RNAs and other biomolecules are essential for maintaining cellular homeostasis. Disruptions in the RNA interactome have been linked to a number of human diseases, implying that these molecular interactions could represent a new family of unexploited therapeutic targets.

Challenges in RNA Interaction Discovery

Despite the growing appreciation of the importance of RNA, discovery and characterization of RNA interactions at the transcriptome level is lagging behind. This is mainly due to the limitations of the existing methods, which include:

• Low precision
• Low throughput
• Low coverage
• Biased analysis
• Complicated protocols involving cumbersome biochemical fractionation or cell-line engineering

With the present technology, many more years may pass before a comprehensive list of their functions, localizations, and interactions can be assembled, considering the immense size and complexity of the human transcriptome and RNA interactome.

Project Objectives

This ERC project aims to establish a simple, versatile, and low-cost technology based on photocatalytic proximity-labeling and the biRhoBAST aptamer for deciphering RNA-RNA and RNA-protein interactions with high precision for any given RNA at different resolutions. These resolutions will range from single-molecule to macromolecular complex levels.

Innovative Technology Integration

Owing to its innovative design, this technology will seamlessly integrate with advanced super-resolution RNA imaging techniques. This integration will provide valuable insights into the intricate interaction networks of RNA with high temporal and spatial resolution.

Expected Outcomes

By applying this massively multiplexable technology to numerous biological settings and disease-related RNAs, we will:

1. Expand our understanding of interactomes
2. Uncover new insights into subcellular RNA structures
3. Unravel fundamental molecular mechanisms of RNA diseases

These efforts will lead to the discovery of novel functions for both RNA and proteins, potentially unlocking new therapeutic targets.