Highly specific gene silencing by artificial miRNAs in rice.
Highly specific gene silencing by artificial miRNAs in rice.
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大米中人造miRNA的高度特异性基因沉默。
DOI：
10.1371/journal.pone.0001829
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发表时间：
2008-03-19
期刊：
影响因子：
3.7
通讯作者：
Herve, Philippe
中科院分区：
文献类型：
--
作者：
Warthmann, Norman;Chen, Hao;Ossowski, Stephan;Weigel, Detlef;Herve, Philippe
内源性microRNAs （miRNAs）是植物和动物基因表达的有效负调控因子。人工miRNAs （amiRNAs）是一种针对一个或几个感兴趣的基因而设计的新方法，为植物有效的转录后基因沉默（PTGS）提供了一种新的和高度特异性的方法。我们为栽培水稻（Oryza sativa）的粳型和籼型品系设计了一种基于amirna的策略。利用水稻内源miRNA前体和定制的21mers，我们设计了针对三个不同基因（Pds、Spl11和Eui1/CYP714D1）的amiRNA构建物。当这些amiRNAs在品种Nipponbare（粳稻）和IR64（籼稻）中组成性表达时，目标基因通过amirna引导的转录物切割而下调，导致预期的突变表型。这种效应对靶基因具有高度特异性，转基因基因稳定遗传，并在后代中保持有效。我们的研究结果不仅表明amiRNAs可以有效地触发单子叶作物的基因沉默，而且还表明amiRNAs可以有效地调节现代育种计划中使用的品种的重要农艺性状。我们提供了设计水稻amiRNA结构的所有软件工具和协议，可以很容易地适应其他作物。该方法适用于候选基因验证、不同品种间的功能基因组比较以及农艺性能和营养价值的提高。
Endogenous microRNAs (miRNAs) are potent negative regulators of gene expression in plants and animals. Artificial miRNAs (amiRNAs)–designed to target one or several genes of interest–provide a new and highly specific approach for effective post-transcriptional gene silencing (PTGS) in plants. We devised an amiRNA-based strategy for both japonica and indica type strains of cultivated rice, Oryza sativa. Using an endogenous rice miRNA precursor and customized 21mers, we designed amiRNA constructs targeting three different genes (Pds, Spl11, and Eui1/CYP714D1). Upon constitutive expression of these amiRNAs in the varieties Nipponbare (japonica) and IR64 (indica), the targeted genes are down-regulated by amiRNA-guided cleavage of the transcripts, resulting in the expected mutant phenotypes. The effects are highly specific to the target gene, the transgenes are stably inherited and they remain effective in the progeny. Our results not only show that amiRNAs can efficiently trigger gene silencing in a monocot crop, but also that amiRNAs can effectively modulate agronomically important traits in varieties used in modern breeding programs. We provide all software tools and a protocol for the design of rice amiRNA constructs, which can be easily adapted to other crops. The approach is suited for candidate gene validation, comparative functional genomics between different varieties, and for improvement of agronomic performance and nutritional value.
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影响因子：
14.9
作者：
Ouyang S;Zhu W;Hamilton J;Lin H;Campbell M;Childs K;Thibaud-Nissen F;Malek RL;Lee Y;Zheng L;Orvis J;Haas B;Wortman J;Buell CR
通讯作者：
Buell CR
影响因子：
14.9
作者：
Griffiths-Jones, S;Bateman, A;Eddy, SR
通讯作者：
Eddy, SR
DOI：
10.1073/pnas.0604421103
发表时间：
2006-11-21
影响因子：
11.1
作者：
Leshem, Yehoram;Melamed-Book, Naomi;Levine, Alex
通讯作者：
Levine, Alex
影响因子：
4.9
作者：
Luo, AD;Qian, Q;Chu, CC
通讯作者：
Chu, CC
影响因子：
4.9
作者：
Miki, D;Shimamoto, K
通讯作者：
Shimamoto, K