Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba shi, Chiba 260-0856, Japan.
Department of Physical Chemistry, School of Pharmacy and Pharmaceutical Sciences, Hoshi University, 2-4-41 Ebara, Shinagawa-Ku, Tokyo 142-8501, Japan.
ACS Nano. 2023 Feb 14;17(3):2588-2601. doi: 10.1021/acsnano.2c10501. Epub 2023 Jan 31.
Based on the clinical success of an transcribed mRNA (IVT-mRNA) that is encapsulated in lipid nanoparticles (mRNA-LNPs), there is a growing demand by researchers to test whether their own biological findings might be applicable for use in mRNA-based therapeutics. However, the equipment and/or know-how required for manufacturing such nanoparticles is often inaccessible. To encourage more innovation in mRNA therapeutics, a simple method for preparing mRNA-LNPs is prerequisite. In this study, we report on a method for encapsulating IVT-mRNA into LNPs by rehydrating a Ready-to-Use empty freeze-dried LNP (LNPs(RtoU)) formulation with IVT-mRNA solution followed by heating. The resulting mRNA-LNPs(RtoU) had a similar intraparticle structure compared to the mRNA-LNPs prepared by conventional microfluidic mixing. genome editing, a promising application of these types of mRNA-LNPs, was accomplished using the LNPs(RtoU) containing co-encapsulated Cas9-mRNA and a small guide RNA.
基于封装在脂质纳米颗粒（mRNA-LNPs）中的经转录的 mRNA（IVT-mRNA）的临床成功，研究人员越来越希望测试他们自己的生物学发现是否可适用于基于 mRNA 的治疗。然而，制造这种纳米颗粒所需的设备和/或专门技术通常难以获得。为了鼓励 mRNA 治疗学的更多创新，制备 mRNA-LNPs 的简单方法是前提。在这项研究中，我们报告了一种通过用 IVT-mRNA 溶液重悬即用型冻干 LNPs（LNPs(RtoU)）制剂并加热来将 IVT-mRNA 封装到 LNPs 中的方法。与通过常规微流混合制备的 mRNA-LNPs 相比，所得到的 mRNA-LNPs(RtoU)具有相似的颗粒内结构。使用包含共包封的 Cas9-mRNA 和小向导 RNA 的 LNPs(RtoU) 完成了对这些类型的 mRNA-LNPs 的基因编辑等有前途的应用。
