Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY, USA.
Copenhagen Hepatitis C Program, Department of Infectious Diseases, Copenhagen University Hospital, Hvidovre, Denmark.
Sci Rep. 2017 Aug 15;7(1):8190. doi: 10.1038/s41598-017-07880-8.
With roles in development, cell proliferation and disease, micro-RNA (miRNA) biology is of great importance and a potential therapeutic target. Here we used cross-linking immunoprecipitation (CLIP) and ligation of miRNA-target chimeras on the Argonaute (AGO) protein to globally map miRNA interactions in the cow. The interactome is the deepest reported to date. miRNA targeting principles are consistent with observations in other species, but with expanded pairing rules. Experimental mapping robustly predicted functional miR-17 regulatory sites. From miRNA-specific targeting for >5000 mRNAs we determined gene ontologies (GO). This confirmed repression of genes important for embryonic development and cell cycle progress by the let-7 family, and repression of those involved in cell cycle arrest by the miR-17 family, but also suggested a number of unappreciated miRNA functions. Our results provide a significant resource for understanding of bovine and species-conserved miRNA regulation, and demonstrate the power of experimental methods for establishing comprehensive interaction maps.
miRNA 生物学在发育、细胞增殖和疾病等方面发挥着重要作用，是潜在的治疗靶点。在这里，我们使用交联免疫沉淀（CLIP）和 Argonaute（AGO）蛋白上 miRNA 靶标嵌合体的连接，在牛中进行了 miRNA 相互作用的全局作图。该相互作用组是迄今为止报道的最深的相互作用组。miRNA 的靶向原则与其他物种的观察结果一致，但配对规则有所扩展。实验作图稳健地预测了功能性 miR-17 调控位点。从针对 >5000 个 mRNA 的 miRNA 特异性靶向作用，我们确定了基因本体论（GO）。这证实了 let-7 家族对胚胎发育和细胞周期进展重要基因的抑制作用，以及 miR-17 家族对细胞周期停滞相关基因的抑制作用，但也提示了一些未被认识到的 miRNA 功能。我们的研究结果为理解牛和物种保守的 miRNA 调控提供了重要资源，并证明了实验方法在建立全面的相互作用图谱方面的强大功能。
