Abstract
Efficient gene transfer is a prerequisite for analysing regulation of transfected promoters. We combined the DNA binding property of the cationic polymer polyethylenimine (PEI) and the potent endocytic activity of adenovirus in a PEI–DNA–adenovirus complex which provided efficient plasmid delivery in differentiated cultured cells. We transfected 3T3-F442A adipocytes, C2.7 myocytes and FAO hepatoma cells with a construct containing the simian virus 40 promoter fused to the chloramphenicol acetyltransferase (CAT) gene, using a combination of PEI and 200 p.f.u. per cell of replication-deficient type 5 adenovirus. Resulting CAT activities varied according to the cell type reaching about 0.6, 8 and 38 units/mg protein for respectively 3T3-F442A, FAO and C2.7 cells. Increases in transfection efficiencies were 140- to 300-fold when compared with those obtained with PEI alone. Then we tested physiologically regulated promoters: the phosphoenolpyruvate carboxykinase gene promoter in 3T3-F442A or FAO cells and the hexokinase II gene promoter in C2.7 myocytes. Gene expression was appropriately increased by clofibrate, dexamethasone and insulin for 3T3-F442A, FAO and C2.7 cells, respectively. Thus, the combination of PEI and adenovirus is a simple, efficient, inexpensive and versatile method of gene transfer which is applicable to several differentiated cells and provides a physiologically coherent transgene regulation. We name this method PEI- adenofection.
Similar content being viewed by others
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Meunier-Durmort, C., Grimal, H., Sachs, L. et al. Adenovirus enhancement of polyethylenimine-mediated transfer of regulated genes in differentiated cells. Gene Ther 4, 808–814 (1997). https://doi.org/10.1038/sj.gt.3300450
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300450
- Springer Nature Limited
Keywords
This article is cited by
-
PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process
Cytotechnology (2013)
-
Celastrol enhances AAV1-mediated gene expression in mice adipose tissues
Gene Therapy (2011)
-
Fast and reversible trapping of surface glycine receptors by gephyrin
Nature Neuroscience (2001)
-
Transfection of large plasmids in primary human myoblasts
Gene Therapy (2001)
-
Efficient adventitial gene delivery to rabbit carotid artery with cationic polymer–plasmid complexes
Gene Therapy (1999)