Effective Small RNA Destruction by the Expression of a Short Tandem Target Mimic in Arabidopsis
Effective Small RNA Destruction by the Expression of a Short Tandem Target Mimic in Arabidopsis
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通过在拟南芥中表达短串联靶标模拟物来有效破坏小 RNA
DOI：
10.1105/tpc.111.094144
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发表时间：
2012-02-01
期刊：
影响因子：
11.6
通讯作者：
Tang, Guiliang
中科院分区：
文献类型：
--
作者：
Yan, Jun;Gu, Yiyou;Tang, Guiliang
MicroRNA (miRNA) 和其他内源性小 RNA 在真核生物中充当基因组、转录组和蛋白质组的序列特异性调节因子。小RNA功能的探究需要一种有效、广泛适用的方法来特异性阻断小RNA功能。在这里，我们报告了一种高效技术的开发，该技术针对拟南芥中的特定内源 miRNA 或小干扰 RNA 进行破坏。我们发现，短串联靶标模拟物 (STTM) 的表达由两个模仿小 RNA 靶位点的短序列组成，由经验确定的最佳大小的接头分隔开，可导致小 RNA 降解核酸酶降解靶标小 RNA。该技术的功效通过针对三种 miRNA 和一种内源 siRNA 的 STTM 引发的强烈且特异性的发育缺陷得到了证明。总之，我们开发了一种破坏内源小RNA的有效方法，从而为植物和潜在动物中小RNA分子的功能基因组学提供了强大的工具。
MicroRNAs (miRNAs) and other endogenous small RNAs act as sequence-specific regulators of the genome, transcriptome, and proteome in eukaryotes. The interrogation of small RNA functions requires an effective, widely applicable method to specifically block small RNA function. Here, we report the development of a highly effective technology that targets specific endogenous miRNAs or small interfering RNAs for destruction in Arabidopsis thaliana. We show that the expression of a short tandem target mimic (STTM), which is composed of two short sequences mimicking small RNA target sites, separated by a linker of an empirically determined optimal size, leads to the degradation of targeted small RNAs by small RNA degrading nucleases. The efficacy of the technology was demonstrated by the strong and specific developmental defects triggered by STTMs targeting three miRNAs and an endogenous siRNA. In summary, we developed an effective approach for the destruction of endogenous small RNAs, thereby providing a powerful tool for functional genomics of small RNA molecules in plants and potentially animals.