MicroRNAs(miRNAs)是一类内源性非编码小RNA,可在转录后水平调节基因的表达,在细胞生长、发育、疾病发生等过程中发挥着重要作用.明确miRNAs所调控的靶基因对阐明miRNAs的功能及在各种生命过程和疾病发生机制的角色非常关键.目前,鉴定miRNAs的靶基因的方法主要计算机预测方法和生物学实验方法.前者对miRNA靶基因的寻找作出巨大贡献,但常存在很多假阳性,必须通过生物学实验方法加以验证.后者涉及单靶基因鉴定技术和高通量多靶基因鉴定技术,高通量技术又包括基因芯片分析技术、蛋白质组学分析技术、RNA连接酶介导的cDNA末端扩增技术和生物化学法等.本文主要对这些高通量技术的应用、优劣进行归纳,并对其改进方向予以讨论.
MicroRNAs(miRNAs) are a class of endogenous non-coding RNAs involved in the regulation of gene expression at the post-transcriptional level, miRNAs play pivotal roles in cell growth, development and disease pathogenesis. Therefore, to identify and validate miRNAs target genes regulated is essential to understand the functions of miRNAs in disease, miRNAs bind the complementary sequences of target mRNAs to mediate translational repression or target degradation and gene silencing, both computer-aided predication and biological experimental screening can be used for target identification. The former may produce a large number of miRNA target genes with higher false positive genes to be further excluded by the biological experiments. The latter approaches can be further greatly facilitated with high-throughput multiple target screening assays, such as microarray, proteome analysis, or RNA ligase mediated rapid amplification of cDNA ends. The applications involving these assays were summarized and compared, together with the discussion on the further development of related technologies.