Leuschner Philipp J F, Martinez Javier
IMBA-Institute of Molecular Biotechnology of the Austrian Academy of Sciences, 1030 Vienna, Dr. Bohr-Gasse 3, Austria.
Methods. 2007 Oct;43(2):105-9. doi: 10.1016/j.ymeth.2007.04.005.
MicroRNAs (miRNAs) constitute a novel class of short, non-coding RNAs that play crucial roles as silencers of gene expression during biological processes such as development, growth control, cell death, and differentiation. To ensure correct function, the expression of miRNAs must be tightly regulated, a process that is believed to take place at the promoter level. Regulation of the miRNA processing cascade has only recently been shown to play an important role as well and we envision the discovery of additional factors affecting or modulating miRNA processing and ultimately miRNA expression. The biochemical analysis of such factors requires a robust assay to monitor miRNA processing. Here, we discuss protocols and techniques that were used to investigate how the expression of brain-specific miRNA miR-138 is controlled at the level of precursor-miRNA (pre-miRNA) processing.
微小RNA（miRNA）构成了一类新型的短链非编码RNA，在诸如发育、生长控制、细胞死亡和分化等生物学过程中，作为基因表达的沉默因子发挥着关键作用。为确保正确功能，miRNA的表达必须受到严格调控，这一过程被认为发生在启动子水平。miRNA加工级联反应的调控最近也被证明发挥着重要作用，并且我们设想会发现影响或调节miRNA加工以及最终miRNA表达的其他因子。对此类因子进行生化分析需要一种可靠的检测方法来监测miRNA加工。在此，我们讨论用于研究脑特异性miRNA miR-138的表达如何在前体miRNA（pre-miRNA）加工水平受到控制的实验方案和技术。
