Efficient Silencing of Endogenous MicroRNAs Using Artificial MicroRNAs in Arabidopsis thaliana

作     者：Andrew L. Eamens Claire Agius Nell A. Smith Peter M. Waterhouse Ming-Bo Wang 

作者机构：CSIRO Plant Industry GPO Box 1600 Canberra ACT 2601 Australia School of Molecular Bioscience University of Sydney Sydney NSW 2006 Australia 

出 版 物：《Molecular Plant》 (分子植物（英文版）)

年 卷 期：2011年第4卷第1期

页      面：157-170页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0902[农学-园艺学] 090102[农学-作物遗传育种] 

基　　金：Commonwealth Scientific and Industrial Research Organisation  CSIRO 

主　　题：MicroRNA artificial microRNA RNA silencing. 

摘      要：We report here that the expression of endogenous microRNAs （miRNAs） can be efficiently silenced in Arabi- dopsis thaliana （Arabidopsis） using artificial miRNA （amiRNA） technology. We demonstrate that an amiRNA designed to target a mature miRNA directs silencing against all miRNA family members, whereas an amiRNA designed to target the stem-loop region of a miRNA precursor transcript directs silencing against only the individual family member targeted. Furthermore, our results indicate that amiRNAs targeting both the mature miRNA and stem-loop sequence direct RNA silencing through cleavage of the miRNA precursor transcript, which presumably occurs in the nucleus of a plant cell during the initial stages of miRNA biogenesis. This suggests that small RNA （sRNA）-guided RNA cleavage in plants occurs not only in the cytoplasm, but also in the nucleus. Many plant miRNA gene families have been identified via sequencing and bio- informatic analysis, but, to date, only a small tranche of these have been functionally characterized due to a lack of ef- fective forward or reverse genetic tools. Our findings therefore provide a new and powerful reverse-genetic tool for the analysis of miRNA function in plants.