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<p>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&#xa0;chain-free mRNA vaccines, but also offers insights for incorporating large cargo into other liquid crystals, such as peptides and/or polymer materials.</p>

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Premixing enables loading of long RNA in cubic phase lipid nanoparticles

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年5月30日

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  • 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,
  • Hojun Kim

摘要

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.