Evaluation of promoter strength for hepatic gene expression in vivo following adenovirus-mediated gene transfer.
Transgene expression in studies of both gene function and gene therapy may be assisted considerably through the use of transcriptional regulatory elements which permit high-level, and/or tissue-specific gene expression. We have therefore evaluated the transcriptional activities of a series of viral and cellular enhancer/promoter elements, both in vitro and in vivo. The five enhancer/promoter elements showing either high-level or hepatocyte-specific expression following transient transfection into hepatoma cells were incorporated into recombinant adenoviruses expressing human alpha 1-antitrypsin (hAAT) for in vivo studies in the liver of immunodeficient and immunocompetent mice. The human elongation factor 1 alpha gene promoter produced 2 mg/ml serum level of hAAT, which is physiologic in humans and will be therapeutic for patients with AAT deficiency. This and all other enhancer/promoters except that of the CMV-IE gene yielded persistent hAAT expression in SCID mice. These findings demonstrate that adenovirus vectors provide an effective system for studies designed to evaluate enhancer/promoter activities in vivo. Several of the enhancer/promoters examined in this study will have significant utility in adenovirus-mediated gene therapy for alpha 1-antitrypsin deficiency and other genetic disorders.
Guo
Z S
ZS
Department of Cell Biology, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030, USA.
Wang
L H
LH
Eisensmith
R C
RC
Woo
S L
SL
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular; D004279:DNA, Viral / Q000032:analysis; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008113:Liver Neoplasms; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D009928:Organ Specificity; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D014407:Tumor Cells, Cultured; D000515:alpha 1-Antitrypsin / Q000032:analysis / Q000235:genetics
D004279:DNA, Viral; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D012333:RNA, Messenger; D000515:alpha 1-Antitrypsin