European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical SciencesSep 11, 2024

Comparing charged fats for delivering DNA using lipid nanoparticles

mRNA Technology Weekly Brief ↗PubMed ↗DOI ↗

Updated Apr 3, 2026

Abstract

Efficient transgene expression was achieved with a minimum amount of 1 pg DNA per initially plated cells.

Lipid nanoparticles (LNPs) are effective for RNA-based gene delivery and may also be promising for DNA delivery in gene replacement therapies.
The study evaluated different ionizable lipids (DLin-MC3-DMA, SM-102, and ALC-0315) for their impact on DNA expression plasmid delivery.
At an optimized N/P ratio of 6, DNA-LNPs exhibited small, spherical, and monodisperse characteristics with anionic ζ-potential.
Zebrafish studies indicated that selected DNA-LNPs achieved prolonged circulation in blood and reduced clearance by macrophages.
In wildtype mice, DNA-LNPs incorporating ALC-0315 showed superior transfection efficiency compared to other lipid types.

Simplified

Lipid nanoparticles (LNPs) are successfully used for RNA-based gene delivery. In the context of gene replacement therapies, however, delivery of DNA expression plasmids using LNPs as a non-viral vector could be a promising strategy for the induction of longer-lasting effects. Therefore, DNA expression plasmids (3 to 4 kbp) coding for fluorescent markers or luciferase were combined with LNPs. Different clinically used ionizable lipids (DLin-MC3-DMA, SM-102, and ALC-0315) were tested to compare their influence on DNA plasmid delivery. DNA-LNPs were characterized with respect to their colloidal properties (size, polydispersity, ζ-potential, morphology), in vitro performance (cellular uptake, DNA delivery, and gene expression), and in vivo characteristics (biodistribution and luciferase gene expression). At an optimized N/P ratio of 6, spherical, small and monodisperse particles with anionic ζ-potential were obtained. Efficient transgene expression was achieved with a minimum amount of 1 pg DNA per initially plated cells. Zebrafish studies allowed selection of DNA-LNPs, which demonstrated prolonged blood circulation, avoidance of macrophage clearance, and vascular extravasation. Our comparative study demonstrates a high impact of the ionizable lipid type on DNA-LNP performance. Superior transfection efficiency of DNA-LNPs containing the ionizable lipid ALC-0315 was confirmed in wildtype mice.

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Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that could have influenced the work reported in this paper.

PubMed

Comparing charged fats for delivering DNA using lipid nanoparticles

Abstract

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Competing interests

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