Quantifying RNA Degradation with Single-Molecule Nanopore Sensing.

Area of Science:

• Biochemistry
• Molecular Biology
• Nanotechnology

Background:

• RNA's chemical instability poses challenges in diagnostics and therapeutics.
• Gel electrophoresis is the standard for RNA degradation assessment but requires substantial RNA quantities (≥100 ng).
• Sensitive methods are needed for analyzing RNA degradation in valuable samples like mRNA vaccines and viral RNA.

Purpose of the Study:

• To introduce and evaluate solid-state nanopore sensing for assessing RNA degradation.
• To demonstrate the method's sensitivity and applicability to low-abundance RNA samples.
• To compare nanopore sensing with traditional gel electrophoresis for RNA integrity analysis.

Main Methods:

• Utilized solid-state nanopore sensing for single-molecule resolution analysis of RNA degradation.
• Tested the method's performance across a wide range of RNA concentrations.
• Evaluated viral RNA degradation under various experimental conditions.

Main Results:

• Nanopore sensing successfully assessed viral RNA degradation with single-molecule resolution.
• The method is effective with significantly lower RNA amounts (as little as 100 pg) compared to gel electrophoresis.
• Demonstrated utility for analyzing RNA samples unsuitable for gel-based methods due to low concentration or high molecular weight.

Conclusions:

• Solid-state nanopore sensing provides a highly sensitive and quantitative approach for evaluating RNA integrity.
• This method expands the analytical capabilities for RNA samples, particularly those with limited quantity or high molecular weight.
• Nanopore sensing represents a valuable advancement for RNA analysis in diverse research and clinical applications.