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Premixing enables loading of long RNA in cubic phase lipid nanoparticles

Harin Jin, Iji Seo, Jongeon Park, Yunhee Seo, Jae Chul Park, Seunghwan Bang, Joonwoo Rhee, Kwan Hyi Lee, Jahyun Koo, Youngdo Jeong, Ji-Hoon Kim () and Hojun Kim ()
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Harin Jin: University of Science and Technology (UST)
Iji Seo: Korea Institute of Science and Technology (KIST)
Jongeon Park: Korea Institute of Science and Technology (KIST)
Yunhee Seo: Korea Institute of Science and Technology (KIST)
Jae Chul Park: Korea Institute of Science and Technology (KIST)
Seunghwan Bang: University of Science and Technology (UST)
Joonwoo Rhee: Korea Institute of Science and Technology (KIST)
Kwan Hyi Lee: Korea Institute of Science and Technology (KIST)
Jahyun Koo: Korea University
Youngdo Jeong: Korea Institute of Science and Technology (KIST)
Ji-Hoon Kim: Korea Institute of Science and Technology (KIST)
Hojun Kim: University of Science and Technology (UST)

Nature Communications, 2025, vol. 16, issue 1, 1-11

Abstract: Abstract Cubic phase lipid nanoparticles (cubosomes) show promise as carriers for nucleic acids due to their outstanding delivery performance and physical stability. However, both theoretical analysis and experimental results have shown that encapsulating long RNA is almost impossible while maintaining a cubic structure. Here we show successful encapsulation of long RNA (up to 4000 bases) in cubosomes by premixing RNA and lipids before self-assembly. Surprisingly, we discovered that long RNA within cubosomes folded into an isotropic phase, similar to other lipid self-assembly structures encapsulating nucleic acids. The cubosome–long RNA complex maintained excellent delivery efficiency even after 24 days at room temperature, underpinning its exceptional stability. Our premixing strategy not only demonstrates the feasibility of developing cold chain-free mRNA vaccines, but also offers insights for incorporating large cargo into other liquid crystals, such as peptides and/or polymer materials.

Date: 2025
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DOI: 10.1038/s41467-025-60380-6

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