基于实时荧光定量反转录PCR技术的动物miRNA表达分析策略

Real-time Quantitative RT-PCR Analysis of Animal miRNA Expression

在人体内有约60%的蛋白质编码基因受到到miRNA调控。近年来,miRNA参与调控的与动物生长相关的细胞周期、发育、分化和新陈代谢等多种生物学过程也越来越受到关注。目前针对miRNA表达检测的常用方法有很多,其中基于反转录PCR(RT-PCR)的放大检测技术是应用范围最广,实施手段最灵活的一种研究策略。主要包括miRNA的全定量PCR法(miR-Q)、基于茎环引物的实时荧光定量PCR法、Poly(A)加尾性通用反转录法、miRNA表达谱的高通量RT-PCR分析和miRNA扩增谱系(mRAP)法。作者对这几种基于实时荧光定量RT-PCR技术的动物miRNA表达分析策略分别进行了介绍,对其原理、方法和应用范畴进行了概括性总结。

About 60% of human protein-coding genes are regulated by miRNA. These genes produce different proteins, which participant in many biological processes. The functions of these proteins are involved in cell cycle, development, differen- tiation and metabolism. Nowadays, researchers can detect the expressions of miRNA by various approaches. However, the RT-PCR-based amplifying techniques are the most flexible methods, which are applied in extensive fields. The RT-PCR-based amplifying techniques mainly include miR-Q RT-PCR for miRNA quantification, end-point stem-loop Real-time RT-PCR for miRNA quantification, Poly（A）-tailed universal reverse transcription, multiplexing RT-PCR for high-throughput miRNA profiling and miRNA amplification profiling （mRAP）. Here we introduce these important Real-time PCR methods for detecting miRNA, resoectivelv. And the theorv, strategy, application and limitation of each method are summarized synoptically.