Jin, Ha Rin Seo, I Ji Park Jong Eon Seo, Yunhee Park, Jae Chul Bang, Seunghwan Rhee, Joonwoo Lee, Kwan Hyi Koo, Jahyun Jeong, Youngdo Kim, Ji Hoon Kim, Hojun 2025-06-05T05:00:09Z 2025-06-05T05:00:09Z 2025-05-31 2025-05 https://pubs.kist.re.kr/handle/201004/152572 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. English Nature Publishing Group Premixing enables loading of long RNA in cubic phase lipid nanoparticles Article 10.1038/s41467-025-60380-6 1 Nature Communications, v.16, no.1 Nature Communications 16 1 N scie scopus 001499628500008 Multidisciplinary Sciences Science & Technology - Other Topics Article