Synergetic cooperation of microRNAs with transcription factors in iPS cell generation.
Synergetic cooperation of microRNAs with transcription factors in iPS cell generation.
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microRNA 与转录因子在 iPS 细胞生成中的协同作用
DOI：
10.1371/journal.pone.0040849
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Chen J;Wang G;Lu C;Guo X;Hong W;Kang J;Wang J
诱导多能干（iPS）细胞首先是通过成纤维细胞中转录因子（TF）的强制表达而产生的。最近，使用带有或不带有其他转录因子的 miRNA 可以更快速、更有效地生成 iPS 细胞。然而，miRNA 和转录因子在多能性获得和维持中的具体和协同作用仍有待进一步研究。在这里，基于小鼠胚胎成纤维细胞（MEF）、感染Oct3/4、Sox2、Klf4和c-Myc（OSKM）1、2、4或8天的MEF、两种iPS细胞系和ES细胞（代表iPS激活和维持步骤）中的miRNA谱，我们发现两个独特的miRNA组负责iPS生成的不同步骤，并且iPS细胞的miRNA表达谱与iPS细胞的miRNA表达谱非常相似。 ES细胞。此外，我们在上调的miRNA的启动子区域寻找转录因子结合位点，发现上调的miRNA如miR-429-200和miR-17簇直接被外源转录因子激活。 miRNA 或 OSKM 候选靶基因集的 GO 和通路富集提供了 iPS 过程完成过程中 miRNA 和 OSKM 之间的分工和协作的清晰图景。与OSKM调节的通路相比，我们发现miRNA在调节iPS特异性通路中发挥着关键作用，例如粘附连接和Wnt信号通路。此外，我们使用 Dicer 敲低阻断 miRNA 的表达，发现这种处理后 miRNA 的水平降低，并且 iPS 生成的效率显着受到抑制。该研究结合高通量分析、生物统计分析和功能实验，为研究miRNA的重要作用、miRNA及相关信号通路的作用机制以及miRNA在体细胞重编程中的更多应用潜力提供了新思路。
Induced pluripotent stem (iPS) cells were first generated by forced expression of transcription factors (TFs) in fibroblasts. Recently, iPS cells have been generated more rapidly and efficiently using miRNAs with or without other transcription factors. However, the specific and collaborative roles of miRNAs and transcription factors in pluripotency acquisition and maintenance remain to be further investigated. Here, based on the miRNA profiling in mouse embryonic fibroblasts (MEFs), MEFs infected with Oct3/4, Sox2, Klf4 and c-Myc (OSKM) for 1, 2, 4, or 8 day, two iPS cell lines and ES cells, representing iPS activation and maintenance steps, we found that two unique miRNA sets are responsible for different steps of iPS generation, and the miRNA expression profiles of iPS cells are very similar to that of ES cells. Furthermore, we searched for transcription factors binding sites at the promoter regions of up-regulated miRNAs, and found that up-regulated miRNAs such as the miR-429-200 and miR-17 clusters are directly activated by exogenous TFs. The GO and pathway enrichment for candidate target gene sets of miRNAs or OSKM provided a clear picture of division and collaboration between miRNAs and OSKM during completion of the iPS process. Compared with the pathways regulated by OSKM, we found that miRNAs play critical roles in regulating iPS-specific pathways, such as the adherens junction and Wnt signaling pathways. Furthermore, we blocked miRNA expression using Dicer knockdown, and found that the level of miRNAs was decreased following this treatment, and the efficiency of iPS generation was significantly repressed. By combining high-throughput analysis, biostatistical analysis and functional experiments, this study provides new ideas for investigating the important roles of miRNAs, the mechanisms of miRNAs and related signaling pathways, and the potential for many more applications of miRNAs in somatic cell reprogramming.
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影响因子：
3
作者：
Lu M;Shi B;Wang J;Cao Q;Cui Q
通讯作者：
Cui Q
影响因子：
46.9
作者：
通讯作者：
--
影响因子：
64.5
作者：
Marson A;Levine SS;Cole MF;Frampton GM;Brambrink T;Johnstone S;Guenther MG;Johnston WK;Wernig M;Newman J;Calabrese JM;Dennis LM;Volkert TL;Gupta S;Love J;Hannett N;Sharp PA;Bartel DP;Jaenisch R;Young RA
通讯作者：
Young RA
DOI：
10.1073/pnas.0711983105
发表时间：
2008-02-26
影响因子：
11.1
作者：
Lowry, W. E.;Richter, L.;Plath, K.
通讯作者：
Plath, K.
影响因子：
2.8
作者：
Liu, Haiying;Deng, Suo;Guan, Yongmei
通讯作者：
Guan, Yongmei