Conserved enhancer in the serum response factor promoter controls expression during early coronary vasculogenesis.
Conserved enhancer in the serum response factor promoter controls expression during early coronary vasculogenesis.
复制标题
血清反应因子启动子中的保守增强子控制早期冠状血管发生期间的表达。
DOI：
10.1161/01.res.0000125296.14014.17
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Misra,RaviP
中科院分区：
文献类型：
--
作者：
Nelson,TimothyJ;Duncan,StephenA;Misra,RaviP
血清反应因子（SRF）是发育中的小鼠胚胎中中胚层形成所需的转录因子，其对于肌源性分化是重要的，包括值得注意的是，在早期发育期间心外膜器官（PEO）分化成冠状血管细胞。为了确定在小鼠发育早期控制SRF表达的调控序列，我们使用了一种新的转基因方法来研究保守的非编码DNA序列（CNCS）在SRF基因中的作用。使用含有推定SRF调控序列的靶向单拷贝的胚胎干（ES）细胞通过四倍体聚集直接产生转基因胚胎。因为ES细胞衍生的靶向胚胎在遗传上是等同的，除了推定的感兴趣的调控序列，转基因表达的差异可以直接归因于这些序列。使用这种方法，我们确定了一个E-box/Ets包含270-bpcs的SRF启动子中的作用模块，介导的PEO中的表达。含有该模块的报告转基因在产生冠状血管的PEO衍生物中表达，但在PEO衍生的心外膜中不表达。这些结果是第一次报道在体内分析的SRF调控元件，控制在早期发展的表达。使用这种报告模块和这种方法，它应该是可能的开始阐明参与冠状动脉血管祖细胞分化的分子机制，以及确定其他SRF调节元件在哺乳动物发育过程中的重要性。
Serum response factor (SRF) is a transcription factor required for mesoderm formation in the developing mouse embryo that is important for myogenic differentiation, including notably, the differentiation of the proepicardial organ (PEO) into coronary vascular cells during early development. To identify regulatory sequences that control SRF expression during early mouse development, we used a novel transgenic approach to study the role of conserved noncoding DNA sequences (CNCS) in the SRF gene. Embryonic stem (ES) cells containing a targeted single-copy of putative SRF regulatory sequences were used to directly generate transgenic embryos by tetraploid aggregation. Because the ES cell-derived targeted embryos are genetically equivalent, except for the putative regulatory sequence of interest, differences in transgene expression can be attributed directly to these sequences. Using this approach, we identified an E-box/Ets containing 270-bpcis-acting module in the SRF promoter that mediates expression in the PEO. Reporter transgenes containing this module express in derivatives of the PEO that give rise to the coronary vasculature, but do not express in the PEO-derived epicardium. These results are the first reported in vivo analysis of SRF regulatory elements that control expression during early development. Using this reporter module and this approach, it should be possible to begin to elucidate molecular mechanisms involved in the differentiation of coronary vasculature progenitor cells, as well as identify additional SRF regulatory elements important during mammalian development.