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The Human Cytomegalovirus Immediate-Early Promoter is Transcriptionally Active in Undifferentiated Mouse Embryonic Stem Cells

Immunology Group, Paterson Institute for Cancer Research (PICR), Christie Hospital NHS Trust, Manchester, United Kingdom

Key Words. Embryonic stem cells • CMV • Transgene expression • Promoter • EGFP

Chris Ward, Ph.D. Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester, M20 4BX, United Kingdom. Telephone: 44-161-446-3157; Fax: 44-161-446-3109; e-mail: wardcm{at}hotmail.com

It has been reported recently that the cytomegalovirus (CMV) immediate-early promoter is transcriptionally inactive in undifferentiated mouse embryonic stem (ES) cells. This result is surprising, since the CMV promoter is used to express transgenes in a variety of cell lines. We studied the expression of a human CMV-driven enhanced green fluorescent protein (EGFP) reporter gene (pEGFP-N1) in five undifferentiated mouse ES cell lines (BL/6III, D3, E14TG2a, MESC20, and 129) and found EGFP expression in all of these cell lines. Under optimal conditions, between 50%-80% transfection efficiencies could be achieved, and EGFP expression levels were maintained for at least 72 hours. Therefore, the human CMV promoter remains a useful system for transgene expression in undifferentiated ES cells.

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