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首页> 外文期刊>Cell biology international. >CMV promotor activity during ES cell differentiation: potential insight into embryonic stem cell differentiation.
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CMV promotor activity during ES cell differentiation: potential insight into embryonic stem cell differentiation.

机译：ES细胞分化过程中CMV启动子活性：对胚胎干细胞分化的潜在见解。

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The activity of the P(CMV IE) promoter was studied during the differentiation of ES cells into neurons. In order to do this, stable embryonic stem (ES) cell lines that express enhanced green fluorescent protein (EGFP) under the control of P(CMV IE) were created and these ES cells were differentiated by aggregation of cells in the presence of retinoic acid (RA). Based on our observations that the activity of P(CMV IE) was highest in undifferentiated cells, and that cell-cell interaction and addition of RA that lead to enhanced cell proliferation also increased expression from P(CMV IE), we hypothesized that the activity of P(CMV IE) was positively regulated in cycling cells. However, when analysis was done at the single cell level it was found that BrdU label and EGFP expression were not correlated. EGFP expression was found to be down-regulated in many cells that were BrdU positive and conversely there were significant numbers of BrdU negative cells that were EGFP positive. Further, P(CMV IE) activity was not observed in cells that were nestin positive or in differentiated neurons, but P(CMV IE) was active in cells with a fibroblast-like morphology. Finally, several proteins present in undifferentiated ES cells were found to bind to regulatory regions of P(CMV IE). These were absent when cells were aggregated in the presence of RA. The above results have implications for expression of transgenes in ES cells as well as providing new insight into the mechanism of lineage restriction.
机译：在ES细胞分化为神经元的过程中研究了P（CMV IE）启动子的活性。为此，创建了在P（CMV IE）的控制下表达增强的绿色荧光蛋白（EGFP）的稳定的胚胎干（ES）细胞系，并在视黄酸存在下通过聚集细胞来分化这些ES细胞。 （RA）。根据我们的观察，P（CMV IE）的活性在未分化细胞中最高，并且细胞间相互作用和RA的添加导致细胞增殖增强也增加了P（CMV IE）的表达，我们假设该活性P（CMV IE）的表达在周期细胞中受到正调控。然而，当在单细胞水平上进行分析时，发现BrdU标记和EGFP表达不相关。发现在许多BrdU阳性细胞中EGFP表达下调，相反，有大量EGFP阳性BrdU阴性细胞。此外，在巢蛋白阳性或分化的神经元细胞中未观察到P（CMV IE）活性，但在具有成纤维细胞样形态的细胞中P（CMV IE）活性。最后，发现未分化的ES细胞中存在的几种蛋白质与P（CMV IE）的调节区结合。当细胞在RA存在下聚集时，这些不存在。以上结果暗示了转基因在ES细胞中的表达以及对谱系限制机制的新见解。

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