Highly inducible expression from vectors containing multiple GRE's in CHO cells overexpressing the glucocorticoid receptor.
A conditional glucocorticoid-responsive expression vector system is described for highly inducible expression of heterologous genes in mammalian cells. This host-vector system requires high level expression of the glucocorticoid receptor (GR) protein in the host cell and multiple copies of the receptor binding site within the expression vector. Transfection and selection of Chinese hamster ovary cells with expression vectors encoding the rat GR yielded cell lines which express functional receptor at high levels. Insertion of multiple copies of the MMTV enhancer (glucocorticoid responsive element, GRE) into an Adenovirus major late promoter (AdMLP) based expression vector yielded greater than 1000-fold inducible expression by dexamethasone (dex) in transient DNA transfection assays. The induced expression level was 7-fold greater than that obtained with an AdMLP based vector containing an SV40 enhancer, but lacking GRE's. Vectors containing the SV40 enhancer in combination with multiple GRE's exhibited elevated basal expression in the absence of dex, but retained inducibility in both transient assays and after integration and amplification in the CHO genome. This expression system should be of general utility for studying gene regulation and for expressing heterologous genes in a regulatable fashion.
Israel
D I
DI
Genetics Institute, Cambridge, MA 02140.
Kaufman
R J
RJ
D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D003907:Dexamethasone / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D010053:Ovary; D010957:Plasmids; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
D011965:Receptors, Glucocorticoid; D011994:Recombinant Proteins; D003907:Dexamethasone; D015500:Chloramphenicol O-Acetyltransferase