Single-Stranded siRNAs Activate RNAi in Animals
Single-Stranded siRNAs Activate RNAi in Animals
复制标题
DOI：
10.1016/j.cell.2012.08.014
复制
发表时间：
2012-08-31
期刊：
影响因子：
64.5
通讯作者：
Crooke, Stanley T.
中科院分区：
文献类型：
--
作者：
Lima, Walt F.;Prakash, Thazha P.;Crooke, Stanley T.
SiRNAs的治疗作用受到将其输送到组织的复杂制剂的要求。如果能够确定有效的单链RNA，它们将为药理药物提供一条更简单的途径。在这里，我们描述了在没有脂质制剂的动物中沉默基因表达的单链siRNAs(ss-siRNAs)。通过迭代设计，通过确定化学修饰的单链和Argavite 2(AGO2)活性、细胞效力、核酸酶稳定性和药代动力学之间的结构-活性关系来鉴定有效的ss-siRNA。我们发现，乘客链并不是有效基因沉默所必需的。导向链的活动需要AGO2，通过RNAi途径展示作用。SS-siRNA的作用需要5‘-磷酸才能在体内实现活性，我们开发了代谢稳定的5’-(E)-乙烯基膦酸(5‘-VP)，其构象和空间电子性质与天然磷酸相似。识别有效的ss-siRNAs为RNAi治疗提供了另一种选择，并为RNAi机制提供了另一种视角。
The therapeutic utility of siRNAs is limited by the requirement for complex formulations to deliver them to tissues. If potent single-stranded RNAs could be identified, they would provide a simpler path to pharmacological agents. Here, we describe single-stranded siRNAs (ss-siRNAs) that silence gene expression in animals absent lipid formulation. Effective ss-siRNAs were identified by iterative design by determining structure-activity relationships correlating chemically modified single strands and Argonaute 2 (AGO2) activities, potency in cells, nuclease stability, and pharmacokinetics. We find that the passenger strand is not necessary for potent gene silencing. The guide-strand activity requires AGO2, demonstrating action through the RNAi pathway. ss-siRNA action requires a 5' phosphate to achieve activity in vivo, and we developed a metabolically stable 5'-(E)-vinylphosphonate (5'-VP) with conformation and sterioelectronic properties similar to the natural phosphate. Identification of potent ss-siRNAs offers an additional option for RNAi therapeutics and an alternate perspective on RNAi mechanism.