Lipid nanoparticles containing zwitterionic lipids versatilely enhance the efficiency of mRNA delivery

Feb 10, 2026Journal of controlled release : official journal of the Controlled Release Society

Lipid nanoparticles with zwitterionic lipids improve mRNA delivery efficiency

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mRNA Technology on OpenScience ↗PubMed ↗DOI ↗

Abstract

LNPs containing TOT-15 achieved a 5-fold increase in gene expression compared to conventional DSPC-LNPs.

The development of zwitterionic tri-oleoyl-Tris (zTOT) lipids aims to improve mRNA delivery efficiency.
TOT-15-LNPs demonstrated almost 100% gene knockout efficiency without causing toxicity.
Endosomal escape efficiency for TOT-15-LNPs was calculated to be over 9.6%.
The interaction of the TOT-15 system with certain phospholipids promotes membrane fusion and enhances delivery.

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Ionizable lipids enable lipid nanoparticles (LNPs) to deliver nucleic acids, including mRNA, to target cells. However, the efficiency of the endosomal escape of conventional LNPs remains limited to a small percentage. Lipid components that actively interact with net-neutral phospholipids in endosomal membranes and promote membrane fusion potentially enhance the efficiency of endosomal escape and further improve functional mRNA delivery. In this study, we developed a zwitterionic tri-oleoyl-Tris (zTOT) library and used it to enhance mRNA delivery by replacing the widely used helper lipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) in LNPs with zTOT. LNPs containing TOT-15, an example of zTOT, demonstrated a 5-fold higher gene expression compared with that of DSPC-LNPs. TOT-15-LNPs composed of commercially available ionizable lipids showed enhanced gene expression compared with that of DSPC-LNPs. The TOT-15-LNPs also showed almost 100% gene knockout efficiency with no toxicity. The TOT-15 system interacts with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and induces a non-lamellar structure. Pharmacokinetic calculation revealed that endosomal escape efficiency of the TOT-15-LNPs was more than 9.6%. In summary, the results of this study demonstrate that the zTOT system enhances the efficiency of both endosomal escape and functional mRNA delivery.