Duan Shiwei, Wang Yunliang, Wang Hongwei, Wang Shufei, Ji Lindan, Dai Dongjun, Jiang Danjie, Zhang Xiaoxi, Wang Qiang
1. Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, Ningbo, ZJ 315000, China.
2. The Neurology Department of the 148th Hospital of PLA, Zibo, SD 255300, China.
Int J Med Sci. 2014 Oct 11;11(12):1270-4. doi: 10.7150/ijms.9343. eCollection 2014.
MiRNAs are potent regulators of gene expression, and most miRNAs have from several to several thousands of gene targets. Validating the numerous gene targets of a given miRNA remains challenging despite the existence of various tools and databases that predict candidate gene-miRNA pairs. In the present study, we present a high-throughput but flexible method that applies a PCR-based application to simulate the binding of miRNAs to their gene targets. Using hsa-miR-377 as an illustrative example, our method was able to identify 13 potential targets of hsa-miR-377. Moreover, our results include 2 genes (SOD2 and PPM1A) that have already been verified as targets of hsa-miR-377. Our method may provide an alternative way of identifying the gene targets of miRNAs for future research.
微小RNA（miRNA）是基因表达的有效调节因子，大多数miRNA具有从几个到数千个基因靶点。尽管存在各种预测候选基因-miRNA对的工具和数据库，但验证给定miRNA的众多基因靶点仍然具有挑战性。在本研究中，我们提出了一种高通量但灵活的方法，该方法应用基于聚合酶链反应（PCR）的技术来模拟miRNA与其基因靶点的结合。以hsa-miR-377为例，我们的方法能够识别出hsa-miR-377的13个潜在靶点。此外，我们的结果包括2个已被证实为hsa-miR-377靶点的基因（超氧化物歧化酶2（SOD2）和蛋白磷酸酶1A（PPM1A））。我们的方法可能为未来研究鉴定miRNA的基因靶点提供一种替代方法。
