Scherr Michaela, Venturini Letizia, Eder Matthias
Department of Hematology, Hemostasis, Oncology, and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
Methods Mol Biol. 2010;614:175-85. doi: 10.1007/978-1-60761-533-0_12.
Micro (mi)RNAs are highly conserved small regulatory RNAs, which regulate gene expression by hybridization to specific binding sites in the 3'untranslated region (UTR) of many mRNAs. Upon miRNA-guided recruitment of a multiprotein complex, target mRNAs are either degraded or their translation is blocked depending on the complementarity between the miRNAs and their binding sites in target mRNAs. Individual miRNAs have been shown to regulate the expression of hundreds of genes with corresponding miRNA binding sites in the 3'UTR in a dose-dependent manner. Although miRNA-target genes may be predicted by bioinformatic tools, each potential target needs to be confirmed experimentally. We describe here the expression of individual miRNAs or miRNA-specific antagomiRs by lentiviral gene transfer to induce stable gain- and loss-of-function phenotypes. These techniques provide some tools to analyze miRNA function in cell culture or animal models.
微小（mi）RNA是高度保守的小调节RNA，其通过与许多mRNA的3'非翻译区（UTR）中的特定结合位点杂交来调节基因表达。在miRNA引导募集多蛋白复合物后，根据miRNA与其在靶mRNA中的结合位点之间的互补性，靶mRNA要么被降解，要么其翻译被阻断。已表明单个miRNA以剂量依赖性方式调节数百个在3'UTR中具有相应miRNA结合位点的基因的表达。尽管可以通过生物信息学工具预测miRNA靶基因，但每个潜在靶标都需要通过实验进行确认。我们在此描述通过慢病毒基因转移来诱导稳定的功能获得和功能丧失表型，从而实现单个miRNA或miRNA特异性抗miR的表达。这些技术提供了一些工具来分析细胞培养或动物模型中的miRNA功能。
