Department of RNA Therapeutics, Merck Research Laboratories, Merck & Co., Inc., West Point, Pennsylvania, USA.
J Histochem Cytochem. 2013 Jun;61(6):407-20. doi: 10.1369/0022155413484152. Epub 2013 Mar 14.
Small interfering RNA (siRNA) therapeutics have advanced from bench to clinical trials in recent years, along with new tools developed to enable detection of siRNA delivered at the organ, cell, and subcellular levels. Preclinical models of siRNA delivery have benefitted from methodologies such as stem-loop quantitative polymerase chain reaction, histological in situ immunofluorescent staining, endosomal escape assay, and RNA-induced silencing complex loading assay. These technologies have accelerated the detection and optimization of siRNA platforms to overcome the challenges associated with delivering therapeutic oligonucleotides to the cytosol of specific target cells. This review focuses on the methodologies and their application in the biodistribution of siRNA delivered by lipid nanoparticles.
近年来，小干扰 RNA (siRNA) 治疗药物已经从实验室走向临床试验，同时也开发了新的工具来实现对器官、细胞和亚细胞水平递送到的 siRNA 的检测。siRNA 递送的临床前模型受益于一些方法，如茎环定量聚合酶链反应、组织原位免疫荧光染色、内体逃逸测定和 RNA 诱导沉默复合物加载测定。这些技术加速了 siRNA 平台的检测和优化，以克服将治疗性寡核苷酸递送到特定靶细胞胞质溶胶的相关挑战。本综述重点介绍了这些方法及其在脂质纳米颗粒递送到的 siRNA 生物分布中的应用。
