Teotia Sachin, Zhang Dabing, Tang Guiliang
Department of Biological Sciences, Michigan Technological University, Houghton, MI, 49931, USA.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.
Methods Mol Biol. 2017;1654:337-349. doi: 10.1007/978-1-4939-7231-9_25.
Small RNAs, including microRNAs (miRNAs), are abundant in plants and play key roles in controlling plant development and physiology. miRNAs regulate the expression of the target genes involved in key plant processes. Due to functional redundancy among miRNA family members in plants, an ideal approach to silence the expression of all members simultaneously, for their functional characterization, is desirable. Target mimic (TM) was the first approach to achieve this goal. Short tandem target mimic (STTM) is a potent approach complementing TM for silencing miRNAs in plants. STTMs have been successfully used in dicots to block miRNA functions. Here, we describe in detail the protocol for designing STTM construct to block miRNA functions in rice. Such approach can be applied to silence miRNAs in other monocots as well.
包括微小RNA(miRNA)在内的小RNA在植物中大量存在,并在控制植物发育和生理过程中发挥关键作用。miRNA调节参与植物关键过程的靶基因的表达。由于植物miRNA家族成员之间存在功能冗余,因此需要一种理想的方法来同时沉默所有成员的表达,以进行其功能表征。靶标模拟物(TM)是实现这一目标的第一种方法。短串联靶标模拟物(STTM)是一种补充TM的有效方法,用于沉默植物中的miRNA。STTM已成功用于双子叶植物中以阻断miRNA功能。在此,我们详细描述了设计用于阻断水稻中miRNA功能的STTM构建体的方案。这种方法也可应用于沉默其他单子叶植物中的miRNA。