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Stem Cells 2002;20:139-145 www.StemCells.com
© 2002 AlphaMed Press

Analysis of Different Promoter Systems for Efficient Transgene Expression in Mouse Embryonic Stem Cell Lines

Sangmi Chunga,b,c, Therese Anderssona,b,c, Kai-C. Sonntagb,c, Lars Björklundb,c, Ole Isacsonb,c, Kwang-Soo Kima,c

a Molecular Neurobiology Laboratory,
b Neuroregeneration Laboratory, and
c Udall Parkinson's Disease Center, McLean Hospital, Harvard Medical School, Belmont, Massachusetts, USA

Key Words. Embryonic stem cells • Promoter strength • Transgene expression • In vitro differentiation

Kwang-Soo Kim, Ph.D., Molecular Neurobiology Laboratory, MRC216, McLean Hospital, Harvard Medical School, Belmont, Massachusetts 02478, USA. Telephone: 617-855-2024; Fax: 617-855-3479; e-mail: kskim{at}mclean.harvard.edu

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 ß-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.




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