Strategies for RNA Resonance Assignment by ¹³C/¹⁵N- and ¹H-Detected Solid-State NMR Spectroscopy

Area of Science:

• Biophysical Chemistry
• Structural Biology
• Nuclear Magnetic Resonance Spectroscopy

Background:

• Magic angle spinning solid-state NMR (ssNMR) is versatile for non-soluble/non-crystalline biomolecules.
• ssNMR methodology is rapidly advancing, particularly for protein studies.
• RNA studies using ssNMR are limited due to underdeveloped methodologies, especially for resonance assignment.

Purpose of the Study:

• To review recent progress in RNA resonance assignment methods using ssNMR.
• To discuss approaches for secondary structure determination of RNA by ssNMR.
• To identify optimal conditions and future directions for ssNMR studies of RNA, including in RNP complexes.

Main Methods:

• Critical evaluation of conventional 13C- and 15N-detected ssNMR experiments.
• Analysis of novel 1H-detected ssNMR methods for RNA.
• Review of isotope labeling strategies relevant to RNA ssNMR.

Main Results:

• Resonance assignment remains the most critical and limiting step in RNA ssNMR structure determination.
• Both conventional and novel ssNMR techniques offer distinct advantages and limitations for RNA studies.
• Specific experimental regimes are identified as optimal for RNA investigations by ssNMR.

Conclusions:

• Significant progress has been made in ssNMR methods applicable to RNA.
• Further methodological development is crucial to expand ssNMR applications for RNA structure determination.
• Future research should focus on applying advanced ssNMR techniques to large ribonucleoprotein (RNP) complexes.