CELL-EQ: Cellular Equilibrium of RNA through NMR and Structural-Probing

Time period:

1 December 2025 – 30 November 2027

Project leader:

Katja Petzold

Funder:

Swedish Research Council

Type of award:

Project grant

Total funding:

1,000,000 SEK

dispersion Nuclear Magnetic Resonance (RD NMR), we revealed dynamic processes in microRNA and ribosomes and their impact on function. It has become evident that these structural equilibria are sensitive to changes in cellular composition, such as Mg2+ ions, metabolites, or biomacromolecules, and therefore likely represent a mechanism for the regulation of cellular function in response to environmental factors. Hence, the study of structural equilibria within the cell is required. Current atom -resolution structural biology operates under simplified in vitro conditions providing results that are often insufficient to explain cellular function based on structure. Bridging this divide is important to understand RNA regulation in the context of e.g. stress factors and diseases.My objective is to identify factors modulating the RNA structural equilibrium in cells that lead to a change in function. To this end, I will develop methodologies to investigate RNA dynamics in living human cells using NMR and structural probing.Several methods, such as R1ρ RD, DNP-enhanced solid-state NMR and RNA structural probing have sufficiently matured in vitro to permit elucidation of RNA dynamics within human cells. Alongside our advancements in RNA production, stability, and proof-of- concept in-cell NMR, we will adapt methodology to the cell, by increasing sensitivity of RD NMR experiments and by using DNP. We will deconvolve the NMR signal leveraging high-throughput RNA structural probing, applying these techniques to regulatory RNAs, such as microRNA, or ribosomal RNA.  This comprehensive approach will, for the first time, offer glimpses into the structural changes that underpin RNA´s role in cellular functions. This methodology is the linchpin for understanding RNA function and its manipulation for i.e. nanotechnology and drug development.