Glucose responsiveness of a reporter gene transduced into hepatocytic cells using a retroviral vector.
An MMLV-based retroviral vector containing the chloramphenicol acetyl transferase reporter gene under the control of a glucose-dependent internal promoter derived from the L-type pyruvate kinase gene was constructed. After transfection into psi-CRIP packaging cells, clones producing recombinant retrovirus were selected. These retroviruses were used to infect cultured established hepatocytic cells whose endogenous L-type pyruvate kinase gene is transcriptionally regulated by glucose. In the infected cells, the reporter gene was as responsive to glucose as the endogenous L-type pyruvate kinase gene, and the glucose gene activation was time- and concentration-dependent. The possibility to confer a glucose responsiveness on a transgene carried by a retroviral vector provides a powerful tool in the prospect of gene therapy for diabetes mellitus.
Chen
R
R
Institut Cochin de Génétique Moléculaire, Paris, France.
Doiron
B
B
Kahn
A
A
D000818:Animals; D000990:Antithrombin III / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis*; D003001:Cloning, Molecular; D017931:DNA Primers; D004790:Enzyme Induction / Q000187:drug effects; D005814:Genes, Viral; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011770:Pyruvate Kinase / Q000096:biosynthesis; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D012190:Retroviridae; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D017931:DNA Primers; D012367:RNA, Viral; D011994:Recombinant Proteins; D000990:Antithrombin III; D015500:Chloramphenicol O-Acetyltransferase; D011770:Pyruvate Kinase; D005947:Glucose