Yoshinaga Naoto, Naito Mitsuru, Tachihara Yoshihiro, Boonstra Eger, Osada Kensuke, Cabral Horacio, Uchida Satoshi
Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
RIKEN Center for Sustainable Resource Science, Wako 351-0198, Saitama, Japan.
Pharmaceutics. 2021 May 27;13(6):800. doi: 10.3390/pharmaceutics13060800.
Messenger RNA (mRNA) delivery strategies are required to protect biologically fragile mRNA from ribonuclease (RNase) attacks to achieve efficient therapeutic protein expression. To tackle this issue, most mRNA delivery systems have used cationic components, which form electrostatically driven complexes with mRNA and shield encapsulated mRNA strands. However, cationic materials interact with anionic biomacromolecules in physiological environments, which leads to unspecific reactions and toxicities. To circumvent this issue of cation-based approaches, herein, we propose a cation-free delivery strategy by hybridization of PEGylated RNA oligonucleotides with mRNA. The PEG strands on the mRNA sterically and electrostatically shielded the mRNA, improving mRNA nuclease stability 15-fold after serum incubation compared with unhybridized mRNA. Eventually, the PEGylated mRNA induced nearly 20-fold higher efficiency of reporter protein expression than unhybridized mRNA in cultured cells. This study provides a platform to establish a safe and efficient cation-free mRNA delivery system.
信使核糖核酸（mRNA）递送策略需要保护生物脆弱的mRNA免受核糖核酸酶（RNase）攻击，以实现有效的治疗性蛋白质表达。为了解决这个问题，大多数mRNA递送系统都使用了阳离子成分，这些成分与mRNA形成静电驱动的复合物，并屏蔽包裹的mRNA链。然而，阳离子材料在生理环境中与阴离子生物大分子相互作用，这会导致非特异性反应和毒性。为了规避基于阳离子方法的这个问题，在此，我们提出一种通过聚乙二醇化RNA寡核苷酸与mRNA杂交的无阳离子递送策略。mRNA上的聚乙二醇链在空间和静电上屏蔽了mRNA，与未杂交的mRNA相比，血清孵育后mRNA核酸酶稳定性提高了15倍。最终，在培养细胞中，聚乙二醇化mRNA诱导的报告蛋白表达效率比未杂交的mRNA高出近20倍。这项研究提供了一个建立安全有效的无阳离子mRNA递送系统的平台。
