期刊
STEM CELLS
卷 20, 期 2, 页码 139-145出版社
ALPHAMED PRESS
DOI: 10.1634/stemcells.20-2-139
关键词
embryonic stem cells; promoter strength; transgene expression; in vitro differentiation
资金
- NIMH NIH HHS [R01 MH048866, R29 MH048866, MH 48866] Funding Source: Medline
- NINDS NIH HHS [P50 NS 39793, P50 NS039793, R01 NS084869, P50 NS039793-03] Funding Source: Medline
- NATIONAL INSTITUTE OF MENTAL HEALTH [R01MH048866, R29MH048866] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [P50NS039793] Funding Source: NIH RePORTER
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.
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