ACS applied materials & interfacesJul 16, 2026

Making Negatively Charged mRNA Lipid Nanoparticles More Stable Using Breakable Cholesterol Links

mRNA Technology Weekly Brief ↗PubMed ↗DOI ↗

Updated Jul 17, 2026

Abstract

Crosslinked lipid nanoparticles achieved a significant increase in splenic mRNA expression at 12 hours compared to uncrosslinked formulations.

Anionic helper lipids can influence lipid nanoparticle (LNP) expression towards specific organs, such as the spleen.
Weak RNA-lipid interactions during purification can lead to nanoparticle rearrangement and reduced activity.
The use of a cleavable crosslinking strategy stabilized anionic LNPs without altering the main lipid components.
Tuning the lengths of the crosslinker and PEG-diamine improved structural stability of the formulations.
33.2% of CD45+ tdTomato+ cells in the spleen were identified as T cells after treatment with the optimized crosslinked formulation.
Mechanistic analyses indicate that improved transfection efficiency may result from controlled mRNA release and altered intracellular processing.

Simplified

Lipid nanoparticles (LNPs) are the leading platform for mRNA delivery, with their in vivo performance governed by lipid composition and colloidal stability. While anionic helper lipids can bias LNP expression toward the spleen, weak RNA-lipid interactions during purification often induce nanoparticle rearrangement and reduced activity. These stability limitations effectively narrow the accessible formulation design and screening space, leaving large regions of anionic compositional space underexplored. Here, we extend our cleavable crosslinking strategy to stabilize anionic LNPs without replacing the primary lipid constituents of the parent LNP formulation. By tuning the lengths of the cholesterol-derived acid-cleavable crosslinker and PEG-diamine, we achieved balanced structural stability. The optimized crosslinked formulation exhibited a significant increase in splenic mRNA expression at 12 h compared to the uncrosslinked LNPs. Notably, 33.2% of CD45⁺ tdTomato⁺ cells in the spleen were identified as T cells. Mechanistic analyses suggest that controlled mRNA release and altered intracellular processing contribute to the improved transfection efficiency. Together, these findings define a tunable crosslinking window that expands the accessible design landscape for tissue- and cell-specific mRNA delivery.

Full Text

Full text is available at the source.

View Full Text ↗

Making Negatively Charged mRNA Lipid Nanoparticles More Stable Using Breakable Cholesterol Links

Abstract

Full Text

You found one interesting study. We’ll send the next 7.

What Lands in Your Inbox Each Week:

Recent issues from the mrna technology brief