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RNA Stability01:53

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...

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Replacing Helper Lipids With Cationic Lipids Enhances mRNA Lipid Nanoparticles Stability in Solution.

Rui Chen1, Letao Xu1, Nan Zhang1

  • 1School of Chemical Engineering, The University of Adelaide, Adelaide, South Australia, 5000, Australia.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
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Messenger RNA lipid nanoparticles (mRNA-LNPs) require cold storage. Replacing helper phospholipids with DOTAP enhances mRNA-LNP stability in solution, reducing degradation and improving accessibility.

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Area of Science:

  • Biotechnology
  • Nanomedicine
  • Drug Delivery

Background:

  • Messenger RNA lipid nanoparticles (mRNA-LNPs) are crucial for vaccines but degrade rapidly in solution.
  • Current reliance on cold chain logistics limits accessibility and increases costs.
  • Enhancing in-solution stability is vital for broader mRNA-LNP applications.

Purpose of the Study:

  • To investigate the impact of helper phospholipid composition on mRNA-LNP storage stability.
  • To identify alternative lipids that improve the shelf-life of mRNA-LNPs.
  • To understand the structure-stability relationship in mRNA-LNPs.

Main Methods:

  • Ratiometric replacement of DSPC with alternative lipids (DOPS, DOTAP, DOPE) in a commercial mRNA LNP formulation.
  • Stability assessment of modified mRNA-LNPs at various temperatures (42 °C, 22 °C, 4 °C).
  • Microstructure analysis using cryogenic electron microscopy (cryo-EM).

Main Results:

  • Partial substitution with the cationic lipid DOTAP significantly enhanced mRNA-LNP in-solution stability across tested temperatures.
  • DOTAP substitution promoted the formation of intact LNPs and reduced destabilizing "bleb"-like structures.
  • Improved structural integrity correlated with enhanced storage stability.

Conclusions:

  • Helper phospholipid composition critically influences mRNA-LNP storage stability.
  • DOTAP is a promising alternative lipid for improving mRNA-LNP shelf-life.
  • Findings offer strategies to overcome cold chain limitations for mRNA-LNP therapeutics.