Neubacher Saskia, Arenz Christoph
Institute for Chemistry, Humboldt Universität zu Berlin, Berlin, Germany.
Methods Mol Biol. 2014;1095:109-19. doi: 10.1007/978-1-62703-703-7_9.
The ever-increasing number of different miRNAs and their association with a vast number of cellular dysfunctions and diseases have initiated several groups to investigate miRNA maturation, which ultimately leads to down regulation of a target messenger RNA (mRNA) and its downstream product. A rapid, convenient, and reliable assay to detect the Dicer-mediated miRNA-maturation step may facilitate research in this field. Here we describe the in vitro detection of the Dicer-mediated miRNA maturation step using unmodified pre-miRNA and branched rolling circle amplification.
不同miRNA的数量不断增加,且它们与大量细胞功能障碍和疾病相关,这促使多个研究团队对miRNA成熟过程展开研究,该过程最终导致靶信使核糖核酸(mRNA)及其下游产物的表达下调。一种快速、便捷且可靠的检测Dicer介导的miRNA成熟步骤的方法可能会推动该领域的研究。在此,我们描述了使用未修饰的前体miRNA和分支滚环扩增对Dicer介导的miRNA成熟步骤进行体外检测。