基于荧光共振能量转移原理监测小干扰RNA从脂质纳米颗粒中的释放。
Monitoring the siRNA release from lipid nanoparticles based on the fluorescence resonance energy transfer principle.
作者信息
Sun Lei, Zhang Jinfang, Zhou Jing-E, Wang Jing, Wang Zhehao, Luo Shenggen, Wang Yeying, Zhu Shulei, Yang Fan, Tang Jie, Lu Wei, Wang Yiting, Yu Lei, Yan Zhiqiang
机构信息
Institute of Biomedical Engineering and Technology, Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, China.
Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, China.
出版信息
Asian J Pharm Sci. 2023 Jan;18(1):100769. doi: 10.1016/j.ajps.2022.11.003. Epub 2022 Dec 7.
The siRNA-loaded lipid nanoparticles have attracted much attention due to its significant gene silencing effect and successful marketization. However, the distribution and release of siRNA still cannot be effectively monitored. In this study, based on the fluorescence resonance energy transfer (FRET) principle, a fluorescence dye Cy5-modified survivin siRNA was conjugated to nanogolds (Au-DR-siRNA), which were then wrapped with lipid nanoparticles (LNPs) for monitoring the release behaviour of siRNA . The results showed that once Au-DR-siRNA was released from the LNPs and cleaved by the Dicer enzyme to produce free siRNA in cells, the fluorescence of Cy5 would change from quenched state to activated state, showing the location and time of siRNA release. Besides, the LNPs showed a significant antitumor effect by silencing the survivin gene and a CT imaging function superior to iohexol by nanogolds. Therefore, this work provided not only an effective method for monitoring the pharmacokinetic behaviour of LNP-based siRNA, but also a siRNA delivery system for treating and diagnosing tumors.
负载小干扰RNA（siRNA）的脂质纳米颗粒因其显著的基因沉默效应和成功的市场化而备受关注。然而，siRNA的分布和释放仍无法得到有效监测。在本研究中，基于荧光共振能量转移（FRET）原理，将荧光染料Cy5修饰的生存素siRNA与纳米金缀合（Au-DR-siRNA），然后用脂质纳米颗粒（LNP）包裹以监测siRNA的释放行为。结果表明，一旦Au-DR-siRNA从LNP中释放并在细胞中被Dicer酶切割产生游离siRNA，Cy5的荧光将从淬灭状态转变为激活状态，显示出siRNA释放的位置和时间。此外，LNP通过沉默生存素基因显示出显著的抗肿瘤作用，并且具有优于纳米金碘海醇的CT成像功能。因此，这项工作不仅提供了一种监测基于LNP的siRNA药代动力学行为的有效方法，还提供了一种用于治疗和诊断肿瘤的siRNA递送系统。
Zotero 插件