Tomasello Luisa, Cluts Landon, Croce Carlo M
Department of Cancer Biology and Medical Genetics, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA.
Methods Mol Biol. 2019;1970:315-330. doi: 10.1007/978-1-4939-9207-2_17.
MicroRNAs (miRNAs) are small noncoding RNAs of 22-25 nucleotides that control gene expression at the posttranscriptional level through the degradation of mRNAs or translational repression. In the last 15 years, the study of these small molecules helped elucidate their role in the regulation of many cellular processes and the onset and development of several diseases. Therefore, many computational tools based on algorithms for target prediction have been developed to identify potential miRNA-target interactions. The improvement of experimental approaches to more easily and quickly confirm in silico predictions has become essential for the study of these small RNAs and their molecular functions. In this chapter, we summarized the principal steps of one of the most used techniques for the validation of microRNA targets, the Luciferase assay, thus explaining the underlying principles and the procedures to apply it best.
微小RNA（miRNA）是一类长度为22 - 25个核苷酸的小型非编码RNA，它们通过降解mRNA或抑制翻译在转录后水平控制基因表达。在过去的15年里，对这些小分子的研究有助于阐明它们在许多细胞过程调控以及多种疾病的发生和发展中所起的作用。因此，已经开发了许多基于靶标预测算法的计算工具来识别潜在的miRNA - 靶标相互作用。改进实验方法以更轻松、快速地确认计算机模拟预测结果，对于研究这些小RNA及其分子功能变得至关重要。在本章中，我们总结了用于验证微小RNA靶标的最常用技术之一——荧光素酶测定法的主要步骤，从而解释其基本原理以及最佳应用方法。
