TALEN-based knockout library for human microRNAs
TALEN-based knockout library for human microRNAs
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DOI：
10.1038/nsmb.2701
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发表时间：
2013-12-01
影响因子：
16.8
通讯作者：
Kim, V. Narry
中科院分区：
文献类型：
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作者：
Kim, Young-Kook;Wee, Gabbine;Kim, V. Narry
已经开发了各种技术工具来探测microRNA（miRNAs）的功能，但它们的应用受到低效率和特异性的限制。为了克服这些限制，我们使用转录激活因子样效应核酸酶（TALEN）敲除人类miRNA基因。我们为274个miRNA位点设计并制备了540对TALEN文库，重点关注潜在重要的miRNA。敲除过程仅需2-4周，可应用于任何细胞类型。作为一个案例研究，我们产生了两个相关的miRNAs，miR-141和miR-200 c，这属于高度保守的miR-200家族敲除细胞。有趣的是，miR-141和miR-200 c尽管总体上相似，但在很大程度上抑制了不重叠的靶标组，因此表明功能性miRNA-靶标相互作用需要严格的种子配对。我们的研究说明了TALEN技术的潜力，并为miRNA研究提供了有用的资源。
Various technical tools have been developed to probe the functions of microRNAs (miRNAs), yet their application has been limited by low efficacy and specificity. To overcome the limitations, we used transcription activator-like effector nucleases (TALENs) to knock out human miRNA genes. We designed and produced a library of 540 pairs of TALENs for 274 miRNA loci, focusing on potentially important miRNAs. The knockout procedure takes only 2-4 weeks and can be applied to any cell type. As a case study, we generated knockout cells for two related miRNAs, miR-141 and miR-200c, which belong to the highly conserved miR-200 family. Interestingly, miR-141 and miR-200c, despite their overall similarity, suppress largely nonoverlapping groups of targets, thus suggesting that functional miRNA-target interaction requires strict seed-pairing. Our study illustrates the potency of TALEN technology and provides useful resources for miRNA research.