Beijing Institute of Radiation Medicine, Beijing 100850, PR China.
Nucleic Acids Res. 2012 Jan;40(2):884-91. doi: 10.1093/nar/gkr715. Epub 2011 Sep 21.
microRNAs (miRNAs) are a versatile class of non-coding RNAs involved in regulation of various biological processes. miRNA-122 (miR-122) is specifically and abundantly expressed in human liver. In this study, we employed 3'-end biotinylated synthetic miR-122 to identify its targets based on affinity purification. Quantitative RT-PCR analysis of the affinity purified RNAs demonstrated a specific enrichment of several known miR-122 targets such as CAT-1 (also called SLC7A1), ADAM17 and BCL-w. Using microarray analysis of affinity purified RNAs, we also discovered many candidate target genes of miR-122. Among these candidates, we confirmed that protein kinase, interferon-inducible double-stranded RNA-dependent activator (PRKRA), a Dicer-interacting protein, is a direct target gene of miR-122. miRNA quantitative-RT-PCR results indicated that miR-122 and small interfering RNA against PRKRA may facilitate the accumulation of newly synthesized miRNAs but did not detectably affect endogenous miRNAs levels. Our findings will lead to further understanding of multiple functions of this hepato-specific miRNA. We conclude that miR-122 could repress PRKRA expression and facilitate accumulation of newly synthesized miRNAs.
microRNAs (miRNAs) 是一类具有多种功能的非编码 RNA,参与多种生物学过程的调控。miRNA-122 (miR-122) 在人类肝脏中特异性且大量表达。在本研究中,我们使用 3'-端生物素标记的合成 miR-122 进行亲和纯化,基于亲和纯化鉴定其靶标。对亲和纯化 RNA 的定量 RT-PCR 分析表明,几种已知的 miR-122 靶标如 CAT-1(也称为 SLC7A1)、ADAM17 和 BCL-w 特异性富集。通过对亲和纯化 RNA 的微阵列分析,我们还发现了许多 miR-122 的候选靶基因。在这些候选基因中,我们证实蛋白激酶、干扰素诱导的双链 RNA 依赖性激活剂 (PRKRA)、一种 Dicer 相互作用蛋白,是 miR-122 的直接靶基因。miRNA 定量 RT-PCR 结果表明,miR-122 和针对 PRKRA 的小干扰 RNA 可能促进新合成 miRNA 的积累,但对内源性 miRNA 水平没有明显影响。我们的研究结果将进一步了解这种肝脏特异性 miRNA 的多种功能。我们得出结论,miR-122 可以抑制 PRKRA 的表达并促进新合成 miRNA 的积累。