Stable expression of hrGFP by mouse embryonic stem cells: promoter activity in the undifferentiated state and during dopaminergic neural differentiation.
Three promoters, cellular polypeptide chain elongation factor 1 alpha (EF1), cytomegalovirus (CMV), and Rous sarcoma virus (RSV) were examined for stable transgene expression in mouse embryonic stem (ES) cells and their progeny during dopaminergic neural differentiation. In undifferentiated ES cells the EF1 promoter was highly effective, while CMV had moderate activity. After 3 months in culture, expression of humanized renilla green fluorescent protein (hrGFP) was unchanged for the EF1 promoter and decreased for CMV. At the nestin-positive stage of differentiation, hrGFP and nestin were colocalized in about 20% of cells for EF1, in contrast to 80% of cells for the CMV promoter. In tyrosine hydroxylase (TH)-positive neurons neither the EF1 nor CMV promoter were effective. The RSV promoter was inactive in undifferentiated, nestin-positive, and TH-positive cells. Thus, EF1 and CMV are effective promoters for transgene expression in undifferentiated ES cells and nestin-positive neural precursors.
Section on Development and Plasticity, Cellular Neurobiology Research Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland, USA. xzeng@intra.nida.nih.gov
Xianmin
X
Zeng
Jia
J
Chen
Joseph F
JF
Sanchez
Mark
M
Coggiano
Ora
O
Dillon-Carter
Josiah
J
Petersen
William J
WJ
Freed
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D019076:Transgenes
D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D020648:Peptide Elongation Factor 1; D049452:Green Fluorescent Proteins; D004298:Dopamine