Two miRNA Clusters Reveal Alternative Paths in Late-Stage Reprogramming
Two miRNA Clusters Reveal Alternative Paths in Late-Stage Reprogramming
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DOI：
10.1016/j.stem.2014.01.021
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发表时间：
2014-05-01
期刊：
影响因子：
23.9
通讯作者：
Blelloch, Robert
中科院分区：
文献类型：
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作者：
Parchem, Ronald J.;Ye, Julia;Blelloch, Robert
特异性因子如Oct 4、Sox 2和Klf 4（OSK）的异位表达足以将体细胞重编程为诱导多能干细胞（iPSC）。在这项研究中，我们研究了在重编程过程中细胞所采取的路径，通过以下两个多能miRNA簇（mir-290和mir-302）在体内和体外与敲入报告的单个细胞中的转录激活。在胚胎发育和胚胎干细胞分化过程中，所有细胞依次表达mir-290和mir-302。相反，在OSK诱导的重编程过程中，细胞以随机、无序的方式激活miRNA位点。然而，向OSK混合物中添加Sall 4导致基因座激活的一致的反向序列（mir-302，然后mir-290）并增加重编程效率。这些结果表明，在重编程的后期阶段，细胞可以遵循多条路径，并且任何单个细胞的轨迹都受到引入的因素组合的强烈影响。
Ectopic expression of specific factors such as Oct4, Sox2, and Klf4 (OSK) is sufficient to reprogram somatic cells into induced pluripotent stem cells (iPSCs). In this study, we examine the paths taken by cells during the reprogramming process by following the transcriptional activation of two pluripotent miRNA clusters (mir-290 and mir-302) in individual cells in vivo and in vitro with knockin reporters. During embryonic development and embryonic stem cell differentiation, all cells sequentially expressed mir-290 and mir-302. In contrast, during OSK-induced reprogramming, cells activated the miRNA loci in a stochastic, nonordered manner. However, the addition of Sall4 to the OSK cocktail led to a consistent reverse sequence of locus activation (mir-302 then mir-290) and increased reprogramming efficiency. These results demonstrate that cells can follow multiple paths during the late stages of reprogramming, and that the trajectory of any individual cell is strongly influenced by the combination of factors introduced.