Graphene Oxide-Modified Resin for Selective dsRNA Removal from In Vitro-Transcribed mRNA

Area of Science:

• Biotechnology
• Materials Science
• Molecular Biology

Background:

• Messenger RNA (mRNA) is crucial for rapid vaccine development.
• Double-stranded RNA (dsRNA) is an undesired byproduct of in vitro transcription (IVT) mRNA production.
• dsRNA can reduce mRNA translation efficiency and trigger inflammatory responses.

Purpose of the Study:

• To develop a facile method for removing dsRNA from IVT mRNA.
• To improve the purity and efficacy of mRNA therapeutics.
• To create a purification system for large-scale mRNA production.

Main Methods:

• Development of PEGylated polystyrene resins surface-modified with graphene oxide (GO@PEG-PS).
• Utilizing GO@PEG-PS for selective adsorption of single-stranded mRNA in the presence of Mg²⁺.
• Employing EDTA and urea for desorption of purified mRNA.

Main Results:

• GO@PEG-PS resin effectively removed at least 80% of dsRNA from IVT mRNA.
• Approximately 85% of mRNA was recovered after purification.
• Purified mRNA showed enhanced protein translation and reduced interferon-beta secretion.

Conclusions:

• The GO@PEG-PS resin provides an efficient method for dsRNA removal from IVT mRNA.
• This purification technique enhances mRNA functionality and reduces immunogenicity.
• The developed chromatography system is suitable for large-scale mRNA purification.