Institute of Bioengineering and Nanotechnology, Singapore, Singapore.
J Biosci Bioeng. 2010 Jan;109(1):1-8. doi: 10.1016/j.jbiosc.2009.06.017. Epub 2009 Jul 17.
Transient gene expression is one possible approach to manipulate the signaling pathways that control the proliferation and differentiation of human embryonic stem (hES) cells. We tested in hES cells a range of baculoviral vectors with a human elongation factor-1alpha promoter and various viral regulatory elements and observed the most dramatic augmenting effect on the transient expression when the promoter was used together with the human cytomegalovirus immediate-early gene enhancer and the woodchuck hepatitis virus post-transcriptional regulatory elements. This vector provided a 1.6-fold increase in the percentage of transduced cells (up to 72%) over a vector containing the elongation factor-1alpha promoter alone. The effective baculoviral transduction of hES cells did not affect cell proliferation, expression of embryonic stem cell markers and teratoma formation. This new viral vector for temporary transgene expression might become a useful tool for developmental biology studies and biomedical applications of hES cells.
瞬时基因表达是一种可能的方法来操纵控制人胚胎干细胞（hES）细胞增殖和分化的信号通路。我们在 hES 细胞中测试了一系列带有人类延伸因子-1α启动子和各种病毒调节元件的杆状病毒载体，并观察到当启动子与人类巨细胞病毒立即早期基因增强子和土拨鼠肝炎病毒转录后调节元件一起使用时，对瞬时表达的影响最为显著。与仅含有延伸因子-1α启动子的载体相比，该载体使转导细胞的百分比增加了 1.6 倍（高达 72%）。有效的杆状病毒转导 hES 细胞不会影响细胞增殖、胚胎干细胞标记物的表达和畸胎瘤的形成。这种新的瞬时转基因表达的病毒载体可能成为发育生物学研究和 hES 细胞的生物医学应用的有用工具。
