Analysis of different promoter systems for efficient transgene expression in mouse embryonic stem cell lines
Analysis of different promoter systems for efficient transgene expression in mouse embryonic stem cell lines
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DOI：
10.1634/stemcells.20-2-139
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发表时间：
2002-01-01
期刊：
影响因子：
5.2
通讯作者：
Kim, KS
中科院分区：
文献类型：
--
作者：
Chung, SM;Andersson, T;Kim, KS
小鼠胚胎干细胞（ES细胞）来源于植入前胚胎的内细胞团，具有产生身体所有细胞类型的发育能力。结合高效的遗传操作和体外分化程序，ES细胞是用于发育途径的分子分析的有用系统。我们分析和比较了细胞多肽链延伸因子1 α（EF），细胞-病毒杂交（巨细胞病毒[CMV]立即早期增强子融合鸡β-肌动蛋白[CBA]），和病毒CMV启动子系统在两个ES细胞系的转录活性。当瞬时转染时，EF和CBA启动子有力地驱动报告基因表达，而CMV启动子无活性。我们还证明了EF和CBA启动子有效地驱动了细胞发育不同阶段的基因表达：幼稚ES细胞，胚状体（EB）和神经元前体细胞。相反，CMV启动子在ES细胞或EB中不具有转录活性，但一旦ES细胞分化成神经元前体细胞就具有显著的活性。我们的数据表明，个别启动子具有不同的能力，表达报告基因表达的ES和其他类型的细胞测试。
Mouse embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo and have the developmental capacity to generate all cell types of the body. Combined with efficient genetic manipulation and in vitro differentiation procedures, ES cells are a useful system for the molecular analysis of developmental pathways. We analyzed and compared the transcriptional activities of a cellular polypeptide chain elongation factor 1 alpha (EF), a cellular-virus hybrid (eytomegalo-virus [CMV] immediate early enhancer fused to chicken beta-actin [CBA]), and a viral CMV promoter system in two ES cell lines. When transiently transfected, the EF and CBA promoters robustly drove reporter gene expression, while the CMV promoter was inactive. We also demonstrated that the EF and CBA promoters effectively drove gene expression in different stages of cell development: naive ES cells, embryoid bodies (EBs), and neuronal precursor cells. In contrast, the CMV promoter did not have transcriptional activity in either ES cells or EB but had significant activity once ES cells differentiated into neuronal precursors. Our data show that individual promoters have different abilities to express reporter gene expression in the ES and other cell types tested.