Analysis of Different Promoter Systems for Efficient Transgene Expression in Mouse Embryonic Stem Cell Lines
Open Access
- 1 March 2002
- journal article
- research article
- Published by Oxford University Press (OUP) in The International Journal of Cell Cloning
- Vol. 20 (2), 139-145
- https://doi.org/10.1634/stemcells.20-2-139
Abstract
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 (cytomegalo‐virus [CMV] immediate early enhancer fused to chicken β‐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: naïve 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.Keywords
This publication has 30 references indexed in Scilit:
- Adenoviral vector design for high-level transgene expression in primitive human hematopoietic progenitorsGene Therapy, 2000
- Lentiviral Vectors for Enhanced Gene Expression in Human Hematopoietic CellsMolecular Therapy, 2000
- Embryoid Bodies: An In Vitro Model of Mouse EmbryogenesisExperimental Physiology, 2000
- Leukemia Inhibitory Factor–dependent Transcriptional Activation in Embryonic Stem CellsThe Journal of cell biology, 1997
- Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitroMechanisms of Development, 1996
- Derivation of completely cell culture-derived mice from early-passage embryonic stem cells.Proceedings of the National Academy of Sciences, 1993
- A stable cellular marker for the analysis of mouse chimeras: the bacterial chloramphenicol acetyltransferase gene driven by the human elongation factor la promoterDifferentiation, 1991
- Efficient selection for high-expression transfectants with a novel eukaryotic vectorGene, 1991
- Efficient Transfection and Expression of Heterologous Genes in PC12 CellsDNA and Cell Biology, 1990
- Establishment in culture of pluripotential cells from mouse embryosNature, 1981