title: Chimeric transcriptional control units for improved liver-specific transgene expression.
abstract: Hepatocyte-directed delivery of therapeutic genes is a major field of gene therapy. An important issue in this context is the availability of promoters units providing for maximum transcriptional activity and specificity. Although a number of liver-specific promoters and transcriptional control elements have been identified and used for gene delivery, no systematic study has been performed to identify the best suitable combination of known liver-specific promoter and enhancer elements. We now report the results of a comparative investigation addressing this issue. We tested a total of 25 synthetic transcriptional control units consisting of either of the four core promoters from liver-specific genes linked in various combinations and configurations to hepatocyte-specific enhancer elements. These constructs were analyzed for transcriptional activity in different cell types in cell culture and in mouse liver in vivo. The data lead to the clear conclusion that a combination of the alcohol dehydrogenase 6 (ADH6) basal promoter linked to two tandem copies of an apoplipoprotein E enhancer element is the transcriptional control unit of choice for the liver-specific expression of transgenes.
authors.affiliation: Institute of Molecular Biology and Tumor Research, Philipps-University, Emil-Mannkopff-Strasse 2, 35033 Marburg, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Gehrke
authors.forename: Valérie
authors.initials: V
authors.lastname: Jérôme
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D002790:Cholesterol 7-alpha-Hydroxylase / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005164:Factor IX / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D007275:Injections, Intravenous; D008099:Liver / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008819:Mice, Nude; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection; D019076:Transgenes / Q000235:genetics*; D000515:alpha 1-Antitrypsin / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; D000515:alpha 1-Antitrypsin; D005164:Factor IX; D000426:Alcohol Dehydrogenase; D008156:Luciferases; D002790:Cholesterol 7-alpha-Hydroxylase