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微小RNA模拟物的设计与验证指南。

The guideline of the design and validation of MiRNA mimics.

作者信息

Wang Zhiguo

机构信息

Department of Medicine, Montreal Heart Institute, University of Montreal, Montreal, QC, Canada.

出版信息

Methods Mol Biol. 2011;676:211-23. doi: 10.1007/978-1-60761-863-8_15.

Abstract

The miRNA mimic technology (miR-Mimic) is an innovative approach for gene silencing. This approach is to generate nonnatural double-stranded miRNA-like RNA fragments. Such an RNA fragment is designed to have its 5'-end bearing a partially complementary motif to the selected sequence in the 3'UTR unique to the target gene. Once introduced into cells, this RNA fragment, mimicking an endogenous miRNA, can bind specifically to its target gene and produce posttranscriptional repression, more specifically translational inhibition, of the gene. Unlike endogenous miRNAs, miR-Mimics act in a gene-specific fashion. The miR-Mimic approach belongs to the "miRNA-targeting" and "miRNA-gain-of-function" strategy and is primarily used as an exogenous tool to study gene function by targeting mRNA through miRNA-like actions in mammalian cells. The technology was developed by my research group (Department of Medicine, Montreal Heart Institute, University of Montreal) in 2007 (Xiao, et al. J Cell Physiol 212:285-292, 2007; Xiao et al. Nat Cell Biol, in review).

摘要

微小RNA模拟技术(miR-Mimic)是一种用于基因沉默的创新方法。该方法是生成非天然的双链微小RNA样RNA片段。这种RNA片段被设计成其5'端带有与靶基因3'UTR中所选序列部分互补的基序。一旦引入细胞,这种模拟内源性微小RNA的RNA片段就能特异性结合其靶基因,并对该基因产生转录后抑制,更具体地说是翻译抑制。与内源性微小RNA不同,miR-Mimics以基因特异性方式起作用。miR-Mimic方法属于“微小RNA靶向”和“微小RNA功能获得”策略,主要用作外源性工具,通过在哺乳动物细胞中以类似微小RNA的作用靶向mRNA来研究基因功能。该技术由我的研究小组(蒙特利尔大学蒙特利尔心脏研究所医学系)于2007年开发(Xiao等人,《细胞生理学杂志》212:285 - 292,2007;Xiao等人,《自然细胞生物学》,正在审核)。

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