Knockout of miR-221 and miR-222 reveals common and specific targets for paralogous miRNAs
Knockout of miR-221 and miR-222 reveals common and specific targets for paralogous miRNAs
复制标题
DOI：
10.1080/15476286.2016.1269994
复制
发表时间：
2017-01-01
期刊：
影响因子：
4.1
通讯作者：
Kim, Young-Kook
中科院分区：
文献类型：
--
作者：
Jeong, Geon;Lim, Yeong-Hwan;Kim, Young-Kook
MicroRNAs （miRNAs）通过序列特异性结合3非翻译区（UTR）来调节mRNA的表达。mirna的种子序列是靶位点识别的关键决定因素。同源mirna具有相同的种子序列，但在其3个区域不同，已知可以调节大部分重叠的mrna群。然而，目前还没有研究通过适当的实验方法分析同源mirna之间的功能差异。在这项研究中，我们比较了同源mirna miR-221和miR-222的靶标。利用核酸酶介导的基因组工程技术，我们建立了这些mirna的敲除细胞系，并精确分析了目标调控的差异。我们发现miR-221和miR-222抑制先前确定的靶标CDKN1B和CDKN1C存在差异。尽管这两种mirna都抑制CDKN1B，但只有miR-221抑制CDKN1C。通过转录组分析，我们发现几种不同的靶mrna分别受miR-221和miR-222的独立调控，尽管大量mrna对miR-221和miR-222有共同反应。本研究首次比较了同源mirna对mRNA的调控，说明具有相同种子序列的同源mirna在靶调控上也存在差异。
MicroRNAs (miRNAs) regulate the expression of mRNA through sequence-specific binding of the 3 untranslated region (UTR). The seed sequence of miRNAs is the key determinant for target site recognition. Paralogous miRNAs, which share the same seed sequences but differ in their 3 regions, are known to regulate largely overlapping groups of mRNAs. However, no study has analyzed functional differences between paralogous miRNAs with proper experimental methods. In this study, we compared the targets of paralogous miRNAs, miR-221 and miR-222. Using a nuclease-mediated genome engineering technique, we established knockout cell lines for these miRNAs, and precisely analyzed differences in target regulation. We found that miR-221 and miR-222 suppress the previously identified targets, CDKN1B and CDKN1C, differentially. Whereas both miRNAs suppressed CDKN1B, only miR-221 suppressed CDKN1C. From transcriptome analyses, we found that several different target mRNAs were regulated by each of miR-221 and miR-222 independently, although a large number of mRNAs responded commonly to miR-221 and miR-222. This is the first study to compare the mRNA regulations by paralogous miRNAs and illustrate that paralogous miRNAs with the same seed sequence also have difference in target regulation.