Zeng Xianmin, Chen Jia, Sanchez Joseph F, Coggiano Mark, Dillon-Carter Ora, Petersen Josiah, Freed William J
Section on Development and Plasticity, Cellular Neurobiology Research Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland, USA.
Stem Cells. 2003;21(6):647-53. doi: 10.1634/stemcells.21-6-647.
Three promoters, cellular polypeptide chain elongation factor 1 alpha (EF1), cytomegalovirus (CMV), and Rous sarcoma virus (RSV) were examined for stable transgene expression in mouse embryonic stem (ES) cells and their progeny during dopaminergic neural differentiation. In undifferentiated ES cells the EF1 promoter was highly effective, while CMV had moderate activity. After 3 months in culture, expression of humanized renilla green fluorescent protein (hrGFP) was unchanged for the EF1 promoter and decreased for CMV. At the nestin-positive stage of differentiation, hrGFP and nestin were colocalized in about 20% of cells for EF1, in contrast to 80% of cells for the CMV promoter. In tyrosine hydroxylase (TH)-positive neurons neither the EF1 nor CMV promoter were effective. The RSV promoter was inactive in undifferentiated, nestin-positive, and TH-positive cells. Thus, EF1 and CMV are effective promoters for transgene expression in undifferentiated ES cells and nestin-positive neural precursors.
研究了三种启动子，即细胞多肽链延伸因子1α（EF1）、巨细胞病毒（CMV）和劳氏肉瘤病毒（RSV），以检测其在小鼠胚胎干细胞（ES细胞）及其多巴胺能神经分化后代中稳定转基因表达的情况。在未分化的ES细胞中，EF1启动子非常有效，而CMV启动子活性中等。培养3个月后，EF1启动子驱动的人源化海肾绿色荧光蛋白（hrGFP）表达无变化，而CMV启动子驱动的hrGFP表达下降。在巢蛋白阳性分化阶段，EF1启动子驱动的hrGFP与巢蛋白在约20%的细胞中共定位，而CMV启动子驱动的hrGFP与巢蛋白在80%的细胞中共定位。在酪氨酸羟化酶（TH）阳性神经元中，EF1和CMV启动子均无效。RSV启动子在未分化细胞、巢蛋白阳性细胞和TH阳性细胞中均无活性。因此，EF1和CMV是未分化ES细胞和巢蛋白阳性神经前体细胞中转基因表达的有效启动子。
