Embryonic stem cell-specific microRNAs promote induced pluripotency. Nat Biotechnol

The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, California, USA.
Nature Biotechnology (Impact Factor: 41.51). 05/2009; 27(5):459-61. DOI: 10.1038/nbt.1535
Source: PubMed


This report demonstrates that introduction of microRNAs (miRNAs) specific to embryonic stem cells enhances the production of mouse induced pluripotent stem (iPS) cells. The miRNAs miR-291-3p, miR-294 and miR-295 increase the efficiency of reprogramming by Oct4, Sox2 and Klf4, but not by these factors plus cMyc. cMyc binds the promoter of the miRNAs, suggesting that they are downstream effectors of cMyc during reprogramming. However, unlike cMyc, the miRNAs induce a homogeneous population of iPS cell colonies.

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Available from: Robert Judson, Oct 01, 2015
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    • "In addition to the miRNAs generated from the miR-302/367 and 371–373 clusters, the enriched miRNAs in hESCs also contain miRNAs of the miR-17 and miR-515 families (Stadler et al., 2010). The miRNAs expressed from the miR-290–295 cluster can facilitate G1 → S transition (Wang et al., 2008) and promote induced pluripotency (Judson et al., 2009), while miRNAs co-expressed from the miR-302/367 cluster are sufficient to reprogram fibroblasts into iPSCs (Anokye-Danso et al., 2011). During the ESC differentiation process, the ESC-specific miRNAs were all expressed at high levels in the first and second stages but were expressed at lower levels in the last two stages (Table S3). "
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    ABSTRACT: Embryonic stem cells (ESCs) and induced pluripotent stem cells can be induced to differentiate into retinal pigment epithelium (RPE). miRNAs have been characterized and found playing important roles in the differentiation process of ESCs, but their length and sequence heterogeneity (isomiRs), and their non-canonical forms of miRNAs are underestimated or ignored. In this report, we found some non-canonical miRNAs (dominant isomiRs) in all differentiation stages, and 27 statistically significant editing sites were identified in 24 different miRNAs. Morever, we found marked major-to-minor arm-switching events in 14 pre-miRNAs during the hESC to RPE cell differentiation phases. Our study for the first time reports exploring the variability of miRNA expression during the differentiation of hESCs into RPE cells and the results show that miRNA variability is a ubiquitous phenomenon in the ESC differentiation. Copyright © 2015. Published by Elsevier B.V.
    Gene 05/2015; 569(2). DOI:10.1016/j.gene.2015.05.060 · 2.14 Impact Factor
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    • "Small RNAs are usually generated from noncoding regions of gene transcripts and function to suppress gene expression by translational repression and mRNA degradation (Ambros 2004; Chu and Rana 2006, 2007; Rana 2007; Djuranovic et al. 2011; Huntzinger and Izaurralde 2011). Recent work indicates that ES-specific microRNAs can enhance iPSC induction (Judson et al. 2009) and, specifically, that the hES miR-302 can antagonize the senescence response induced by four-factor expression in human fibroblasts (Banito et al. 2009). In addition, our recent findings suggest that the microRNA biogenesis machinery may be required for efficient reprogramming (Li et al. 2011), and microRNAs induced by OSKM are known to regulate several key pathways affecting reprogramming efficiency , including cell cycle control, the p53 pathway, TGFβ signaling, and MET (Choi et al. 2011; Li et al. 2011; Liao et al. 2011; Subramanyam et al. 2011; Yang et al. 2011a). "
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    ABSTRACT: Somatic cells can be reprogrammed to reach an embryonic stem cell-like state by overexpression of defined factors. Recent studies have greatly improved the efficiency of the reprogramming process but the underlying mechanisms regulating the transition from a somatic to a pluripotent state are still relatively unknown. MicroRNAs (miRs) are small noncoding RNAs that primarily regulate target gene expression post-transcriptionally. Here we present a systematic and comprehensive study of microRNAs in mouse embryonic fibroblasts (MEFs) during the early stage of cell fate decisions and reprogramming to a pluripotent state, in which significant transcriptional and epigenetic changes occur. One microRNA found to be highly induced during this stage of reprogramming, miR-135b, targeted the expression of extracellular matrix (ECM) genes including Wisp1 and Igfbp5. Wisp1 was shown to be a key regulator of additional ECM genes that serve as barriers to reprogramming. Regulation of Wisp 1 is likely mediated through biglycan, a glycoprotein highly expressed in MEFs that is silenced in reprogrammed cells. Collectively, this report reveals a novel link between microRNA-mediated regulation of ECM formation and somatic cell reprogramming, and demonstrates that microRNAs are powerful tools to dissect the intracellular and extracellular molecular mechanisms of reprogramming.
    RNA 10/2014; 20(12). DOI:10.1261/rna.043745.113 · 4.94 Impact Factor
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    • "One of the primary obstacles to the successful application of iPSCs for medical purposes is their low reprogramming efficiency . Significant effort has been devoted to enhancing induced reprogramming efficiency, including approaches focusing on the use of mRNA (Warren et al., 2010), small molecules (Ichida et al., 2009; Li and Rana, 2012; Maherali and Hochedlinger, 2009; Nichols et al., 2009; Silva et al., 2008; Yang et al., 2011b; Ying et al., 2008; Zhu et al., 2011), and microRNAs (Choi et al., 2011; Judson et al., 2009; Kim et al., 2011; Li and He, 2012; Li et al., 2011; Liao et al., 2011; Lipchina et al., 2011; Melton et al., 2010; Pfaff et al., 2011; Subramanyam et al., 2011; Yang and Rana, 2013; Yang et al., 2011a). However, detailed functional insight into the molecular basis of reprogramming is still lacking. "
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    ABSTRACT: Although transcriptome analysis can uncover the molecular changes that occur during induced reprogramming, the functional requirements for a given factor during stepwise cell-fate transitions are left unclear. Here, we used a genome-wide RNAi screen and performed integrated transcriptome analysis to identify key genes and cellular events required at the transition steps in reprogramming. Genes associated with cell signaling pathways (e.g., Itpr1, Itpr2, and Pdia3) constitute the major regulatory networks before cells acquire pluripotency. Activation of a specific gene set (e.g., Utf1 or Tdgf1) is important for mature induced pluripotent stem cell formation. Strikingly, a major proportion of RNAi targets (∼53% to 70%) includes genes whose expression levels are unchanged during reprogramming. Among these non-differentially expressed genes, Dmbx1, Hnf4g, Nobox, and Asb4 are important, whereas Nfe2, Cdkn2aip, Msx3, Dbx1, Lzts1, Gtf2i, and Ankrd22 are roadblocks to reprogramming. Together, our results provide a wealth of information about gene functions required at transition steps during reprogramming.
    Cell Reports 07/2014; 8(2). DOI:10.1016/j.celrep.2014.07.002 · 8.36 Impact Factor
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