Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany.
Plant J. 2012 May;70(3):541-7. doi: 10.1111/j.1365-313X.2011.04896.x. Epub 2012 Feb 14.
Gene silencing is an important tool in the study of gene function. Virus-induced gene silencing (VIGS) and hairpin RNA interference (hpRNAi), both of which rely on small interfering RNAs, together with artificial microRNAs (amiRNA), are amongst the most popular methods for reduction of gene activity in plants. However, all three approaches have limitations. Here, we introduce miRNA-induced gene silencing (MIGS). This method exploits a special 22-nucleotide miRNA of Arabidopsis thaliana, miR173, which can trigger production of another class of small RNAs called trans-acting small interfering RNAs (tasiRNAs). We show that fusion of gene fragments to an upstream miR173 target site is sufficient for effective silencing of the corresponding endogenous gene. MIGS can be reliably used for the knockdown of a single gene or of multiple unrelated genes. In addition, we show that MIGS can be applied to other species by co-expression of miR173.
基因沉默是研究基因功能的重要工具。病毒诱导的基因沉默 (VIGS) 和发夹 RNA 干扰 (hpRNAi) 都依赖于小干扰 RNA，以及人工 microRNA (amiRNA)，是植物中降低基因活性最常用的方法之一。然而，这三种方法都有其局限性。在这里，我们介绍了 miRNA 诱导的基因沉默 (MIGS)。该方法利用了拟南芥特殊的 22 个核苷酸 miRNA，miR173，可以触发另一类称为反式作用小干扰 RNA (tasiRNA) 的小 RNA 的产生。我们证明，将基因片段融合到上游 miR173 靶位点足以有效沉默相应的内源性基因。MIGS 可可靠地用于单个基因或多个不相关基因的敲低。此外，我们还通过共表达 miR173 证明了 MIGS 可以应用于其他物种。
