Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3
Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3
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DOI：
10.1186/1471-213x-8-57
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发表时间：
2008-05-23
影响因子：
--
通讯作者：
Buerki, Kurt
中科院分区：
文献类型：
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作者：
Cinelli, Paolo;Casanova, Elisa A.;Buerki, Kurt
背景：转录因子STAT 3是LIF信号级联的下游靶点。LIF信号传导或激活足以维持胚胎干细胞（ES）处于未分化和多能状态。为了进一步研究STAT 3在建立ES细胞中的重要性，我们在第一步中从表达条件性他莫昔芬依赖性STAT 3-MER融合蛋白的转基因FVB小鼠中获得稳定的多能胚胎干细胞。在第二步中，STAT 3-MER过表达细胞被用来确定STAT 3通路相关基因的表达谱，以确定新的关键球员参与维持多能性在ES cells.Results：转基因STAT 3-MER囊胚产生多能生殖系的ES细胞在高频率的LIF的情况下，当建立在含他莫昔芬的培养基。他莫昔芬诱导的转基因FVB ES细胞系的表达谱显示，与野生型细胞相比，一组26个基因显著上调或下调。四个上调基因（己糖激酶II、Lefty 2、Prame 17、PP 1 rs 15 B）的表达显示限于胚泡的内细胞团（ICM）。这些差异表达的基因代表了维持ES细胞多能性的潜在候选基因。我们最终在ES细胞中过表达了两个候选基因Pem/Rhox 5和Prame 17，并证明了它们的过表达足以维持ES细胞标志物的表达以及在缺乏LIF的情况下多能ES细胞的典型形态。STAT 3-过表达胚泡内细胞团中的MER促进ES细胞的建立，并诱导参与维持胚胎发育的潜在候选基因的上调。多能性。其中两个，Pem/Rhox 5和Prame 17，当在ES细胞中过表达时，能够以LIF非依赖性方式将胚胎干细胞维持在多能状态，如STAT 3或Nanog。
Background: The transcription factor STAT3 is a downstream target of the LIF signalling cascade. LIF signalling or activation is sufficient to maintain embryonic stem (ES) cells in an undifferentiated and pluripotent state. To further investigate the importance of STAT3 in the establishment of ES cells we have in a first step derived stable pluripotent embryonic stem cells from transgenic FVB mice expressing a conditional tamoxifen dependent STAT3-MER fusion protein. In a second step, STAT3-MER overexpressing cells were used to identify STAT3 pathway-related genes by expression profiling in order to identify new key-players involved in maintenance of pluripotency in ES cells.Results: Transgenic STAT3-MER blastocysts yielded pluripotent germline-competent ES cells at a high frequency in the absence of LIF when established in tamoxifen-containing medium. Expression profiling of tamoxifen-induced transgenic FVB ES cell lines revealed a set of 26 genes that were markedly up- or down-regulated when compared with wild type cells. The expression of four of the up- regulated genes (Hexokinase II, Lefty2, Prame17, PP1rs15B) was shown to be restricted to the inner cell mass (ICM) of the blastocysts. These differentially expressed genes represent potential candidates for the maintenance of pluripotency of ES cells. We finally overexpressed two candidate genes, Pem/Rhox5 and Prame17, in ES cells and demonstrated that their overexpression is sufficient for the maintenance of expression of ES cell markers as well as of the typical morphology of pluripotent ES cells in absence of LIF.Conclusion: Overexpression of STAT3-MER in the inner cell mass of blastocyst facilitates the establishment of ES cells and induces the upregulation of potential candidate genes involved in the maintenance of pluripotency. Two of them, Pem/Rhox5 and Prame17, when overexpressed in ES cells are able to maintain the embryonic stem cells in a pluripotent state in a LIF independent manner as STAT3 or Nanog.