1] The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, Program in Biomedical Sciences, University of California San Francisco, San Francisco, California, USA. [2] Department of Urology, University of California San Francisco, San Francisco, California, USA.
Nat Struct Mol Biol. 2013 Oct;20(10):1227-35. doi: 10.1038/nsmb.2665. Epub 2013 Sep 15.
Individual microRNAs (miRNAs) can target hundreds of mRNAs forming networks of presumably cooperating genes. To test this presumption, we functionally screened miRNAs and their targets in the context of dedifferentiation of mouse fibroblasts to induced pluripotent stem cells (iPSCs). Along with the miR-302-miR-294 family, the miR-181 family arose as a previously unidentified enhancer of the initiation phase of reprogramming. Endogenous miR-181 miRNAs were transiently elevated with the introduction of Pou5f1 (also known as Oct4), Sox2 and Klf4 (referred to as OSK), and miR-181 inhibition diminished iPSC colony formation. We tested the functional contribution of 114 individual targets of the two families, revealing 25 genes that normally suppress initiation. Coinhibition of targets cooperatively promoted both the frequency and kinetics of OSK-induced reprogramming. These data establish two of the largest functionally defined networks of miRNA-mRNA interactions and reveal previously unidentified relationships among genes that act together to suppress early stages of reprogramming.
单个 microRNAs (miRNAs) 可以靶向数百个 mRNA，形成可能协同作用的基因网络。为了验证这一假设，我们在小鼠成纤维细胞去分化为诱导多能干细胞 (iPSC) 的过程中，对 miRNAs 及其靶标进行了功能筛选。miR-302-miR-294 家族和 miR-181 家族一起，作为重新编程起始阶段的增强子而被首次鉴定。内源性 miR-181 miRNAs 在引入 Pou5f1（也称为 Oct4）、Sox2 和 Klf4（称为 OSK）时短暂升高，miR-181 抑制减少了 iPSC 集落形成。我们测试了这两个家族的 114 个单个靶标的功能贡献，揭示了 25 个正常抑制起始的基因。两个家族的靶基因共同抑制可协同促进 OSK 诱导的重编程的频率和动力学。这些数据建立了两个最大的 miRNA-mRNA 相互作用的功能定义网络，并揭示了以前未知的共同作用以抑制重编程早期阶段的基因之间的关系。
