Persistent expression of foreign genes in cultured hepatocytes: expression vectors.
We have optimized a liposome-based transfection method that mediated highly efficient stable expression of foreign genes in hepatocytes. Moreover, we have observed that the metallothionein 1 promoter in the bovine papilloma virus-based expression vector drove the highest expression of foreign genes in hepatocytes as compared with the cytomegalovirus and the human polypeptide chain elongation factor 1alpha (EF-1alpha) promoters in the pcDNA 3-based expression vector. The cytomegalovirus promoter failed to yield significant expression in these cells. Furthermore, expression of foreign genes persisted up to at least 15 passages when expression was under the control of either the EF-1alpha or the metallothionein 1 promoter. Thus, these two promoters led to comparable stability of foreign genes in hepatocytes, with the metallothionein 1 promoter yielding a higher level of expression of foreign genes in these cells.
Najjar
S M
SM
Department of Pharmacology and Therapeutics, Medical College of Ohio, 3035 Arlington Avenue, Toledo, OH 43614, USA. snajjar@mco.edu
Lewis
R E
RE
D000818:Animals; D052688:Bovine papillomavirus 1 / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002999:Clone Cells / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D006913:Hydroxysteroid Dehydrogenases; D008081:Liposomes / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008562:Membrane Glycoproteins; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection / Q000379:methods*; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
D002352:Carrier Proteins; D006913:Hydroxysteroid Dehydrogenases; D008081:Liposomes; D008562:Membrane Glycoproteins; D008668:Metallothionein; D009687:Nuclear Proteins; D014157:Transcription Factors; C037643:bile acid binding proteins; C435496:AKR1C2 protein, human; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)