A virus-based miRNA suppression ( VbMS) system for miRNA loss-of-function analysis in plants

文献类型： 外文期刊

第一作者： Yan, Fei

作者： Yan, Fei;Guo, Weixia;Lu, Yuwen;Peng, Jiejun;Zheng, Hongying;Lin, Lin;Chen, Jianping;Wu, Gentu;Chen, Jianping

作者机构：

关键词： Loss-of-function;MicroRNA;MiRNA sponge;Target mimic;Virus

期刊名称：BIOTECHNOLOGY JOURNAL （ 影响因子：4.677； 五年影响因子：4.596 ）

ISSN： 1860-6768

年卷期： 2014 年 9 卷 5 期

页码：

收录情况： SCI

摘要： MicroRNAs (miRNAs) play key roles in plant development and defense against pathogens. To establish the function of individual miRNAs, gain-of-function analysis is usually done by overexpressing a specific miRNA in transgenic plants and has proved very effective. Loss-of-function analysis by the target mimic method is now also increasingly being used. The mimics expressed in the transgenic plants sequester a specific miRNA and lead to changed phenotypes that elucidate miRNA function. However, it takes time to obtain the transgenic plants. To avoid using transgenic plants, we have developed a virus-based miRNA suppression system (VbMS) based on a Tobacco rattle virus vector. The target mimic sequences of miR156, miR319, or miR164 were introduced into the viral genomic RNA, which was then inoculated to Arabidopsis thaliana plants. The resulting phenotypes were consistent with previous reports from transgenic plants, and the expression of targets of the miRNAs was also increased showing that the activity of the miRNAs had been inhibited. VbMS developed here is validated for loss-of-function analysis of miRNA in plants. Moreover, since only simple agroinfiltration rather than transformation is needed, VbMS is suitable for large-scale approaches to miRNA function analysis in plants.

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