de Felippes Felipe Fenselau
Department of Biology, Swiss Federal Institute of Technology (ETH), Zurich, Switzerland,
Methods Mol Biol. 2013;942:379-87. doi: 10.1007/978-1-62703-119-6_20.
In plants, some microRNAs (miRNAs) can trigger the production of secondary small interfering RNAs (siRNAs) from their targets. miRNA-induced gene silencing (MIGS) exploits this unique feature to efficiently downregulate gene expression. The simple flanking of a sequence of interest with the target site for the miR173 (an miRNA able to trigger transitivity) is sufficient to start the production of secondary siRNAs and, consequently, silencing of the target gene. This technique can be easily adapted to promote gene silencing of more than one gene, even with those that share no sequence similarities. This chapter describes the necessary steps for designing and implementing the use of MIGS in plants.
在植物中，一些微小RNA（miRNA）能够促使其靶标产生次级小干扰RNA（siRNA）。miRNA诱导的基因沉默（MIGS）利用这一独特特性来有效下调基因表达。将感兴趣的序列简单地侧翼连接上miR173（一种能够引发传递性的miRNA）的靶位点，就足以启动次级siRNA的产生，进而使靶基因沉默。这项技术可以很容易地进行调整，以促进多个基因的沉默，即使这些基因没有序列相似性。本章描述了在植物中设计和实施MIGS使用的必要步骤。
