Highly specific gene silencing by artificial microRNAs in Arabidopsis
Highly specific gene silencing by artificial microRNAs in Arabidopsis
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DOI：
10.1105/tpc.105.039834
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发表时间：
2006-05-01
期刊：
影响因子：
11.6
通讯作者：
Weigel, D
中科院分区：
文献类型：
--
作者：
Schwab, R;Ossowski, S;Weigel, D
植物microRNA（miRNAs）仅影响少数具有高度序列互补性的靶标，而动物miRNAs通常具有数百个互补性有限的靶标。我们使用人工miRNAs（amiRNAs）来确定天然植物miRNAs的狭窄作用谱是否仅反映了植物miRNAs机制的内在特性，或者是否也是由于过去对具有更广泛特异性的天然miRNAs的选择。amiRNA被设计为靶向单个基因或内源基因组。像天然的miRNAs一样，它们有不同数量的靶点错配。先前确定的天然miRNA的靶标选择参数可以准确地预测amiRNA的直接靶标。根据全基因组表达谱推断，amiRNA的特异性与天然植物miRNA一样高，支持植物miRNA功能需要与靶标进行广泛碱基配对的观点。amiRNA是植物中特异性基因沉默的有效工具，特别是当几个相关但不相同的靶基因需要下调时。我们证明了amiRNA在组织特异性或诱导型启动子下表达时也是活跃的，具有有限的非自主效应。amiRNA的设计原理已经被推广并整合到基于Web的工具（http：//www.example.com）中。wmd.weigelworld.org
Plant microRNAs ( miRNAs) affect only a small number of targets with high sequence complementarity, while animal miRNAs usually have hundreds of targets with limited complementarity. We used artificial miRNAs (amiRNAs) to determine whether the narrow action spectrum of natural plant miRNAs reflects only intrinsic properties of the plant miRNA machinery or whether it is also due to past selection against natural miRNAs with broader specificity. amiRNAs were designed to target individual genes or groups of endogenous genes. Like natural miRNAs, they had varying numbers of target mismatches. Previously determined parameters of target selection for natural miRNAs could accurately predict direct targets of amiRNAs. The specificity of amiRNAs, as deduced from genome-wide expression profiling, was as high as that of natural plant miRNAs, supporting the notion that extensive base pairing with targets is required for plant miRNA function. amiRNAs make an effective tool for specific gene silencing in plants, especially when several related, but not identical, target genes need to be downregulated. We demonstrate that amiRNAs are also active when expressed under tissue-specific or inducible promoters, with limited nonautonomous effects. The design principles for amiRNAs have been generalized and integrated into a Web-based tool (http://wmd.weigelworld.org).