Jung You-Jin, Lim Yong-Beom
Department of Materials Science and Engineering, Yonsei University, Seoul, Republic of Korea.
Methods Mol Biol. 2025;2965:403-416. doi: 10.1007/978-1-0716-4742-4_20.
The mRNA delivery vehicle technology is one of the key factors in developing mRNA-based therapeutics and vaccines. The delivery vehicle must protect the mRNA from degradation, accurately deliver it across body barriers into the target tissue or cell, and properly regulate expression of the protein. Even though the stability and performance of mRNA delivery vehicles is highly dependent on their nanostructure, most vehicles were irregular spherical shapes. In this chapter, we describe a detailed protocol for fabricating nanotubular mRNA delivery vehicles using self-assembling peptide building blocks. We present design principles of the peptide building blocks and formulation strategies of mRNA-peptide complexes. Additionally, we cover methods for characterizing and assessing their thermodynamic stability and intracellular performance. These protocols should help facilitate the development of precisely structure-controlled mRNA delivery vehicles.
mRNA递送载体技术是开发基于mRNA的治疗药物和疫苗的关键因素之一。递送载体必须保护mRNA不被降解，将其准确地递送至身体屏障之外进入靶组织或细胞，并适当地调节蛋白质的表达。尽管mRNA递送载体的稳定性和性能高度依赖于其纳米结构，但大多数载体呈不规则球形。在本章中，我们描述了一种使用自组装肽构建块制造纳米管mRNA递送载体的详细方案。我们介绍了肽构建块的设计原则以及mRNA-肽复合物的配方策略。此外，我们还涵盖了表征和评估其热力学稳定性和细胞内性能的方法。这些方案应有助于促进精确结构控制的mRNA递送载体的开发。
