Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells
Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells
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DOI：
10.1016/j.stem.2008.01.004
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发表时间：
2008-02-01
期刊：
影响因子：
23.9
通讯作者：
Jaenisch, Rudolf
中科院分区：
文献类型：
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作者：
Brambrink, Tobias;Foreman, Ruth;Jaenisch, Rudolf
通过反转录病毒转导Oct 4、Sox 2、Klf 4和c-Myc，可以在分化的鼠和人细胞中诱导多能性。我们已经设计了一种重编程策略，其中这四个转录因子从强力霉素（dox）诱导的慢病毒载体表达。使用这些诱导型构建体，我们从小鼠胚胎成纤维细胞（MEFs）中获得了诱导多能干细胞（PS），并发现转基因沉默是正常细胞分化的先决条件。我们分析了小鼠PS细胞衍生过程中已知多能性标记物激活的时间，并观察到碱性磷酸酶（AP）首先被激活，然后是阶段特异性胚胎抗原1（SSEA 1）。Nanog和内源性Oct 4基因的表达，标志着完全重编程的细胞，仅在该过程的后期观察到。重要的是，病毒转导的cDNA需要表达至少12天以产生PS细胞。我们的研究结果是一个步骤，了解一些分子事件管理表观遗传重编程。
Pluripotency can be induced in differentiated murine and human cells by retroviral transduction of Oct4, Sox2, Klf4, and c-Myc. We have devised a reprogramming strategy in which these four transcription factors are expressed from doxycycline (dox)-inducible lentiviral vectors. Using these inducible constructs, we derived induced pluripotent stem (PS) cells from mouse embryonic fibroblasts (MEFs) and found that transgene silencing is a prerequisite for normal cell differentiation. We have analyzed the timing of known pluripotency marker activation during mouse PS cell derivation and observed that alkaline phosphatase (AP) was activated first, followed by stage-specific embryonic antigen 1 (SSEA1). Expression of Nanog and the endogenous Oct4 gene, marking fully reprogrammed cells, was only observed late in the process. Importantly, the virally transduced cDNAs needed to be expressed for at least 12 days in order to generate PS cells. Our results are a step toward understanding some of the molecular events governing epigenetic reprogramming.