title: Novel, high expressing and antibiotic-controlled plasmid vectors designed for use in gene therapy.
abstract: The promise of effective gene therapy can only be accomplished by high-level expression and regulatable delivery of gene products. To achieve this end, a eukaryotic expression plasmid was modified to make transcription dependent on a tetracycline(Tc)-regulated chimeric transactivator. Mouse muscle injected with this two plasmid cis/trans control system expressed reporter proteins at levels five- to 10-fold greater than the cytomegalovirus immediate-early promoter-controlled parental plasmid. Tetracycline could be useful to either repress or activate transactivator-controlled expression based on the position of the tetO control sequences within the reporter plasmid. Finally, a prototype single plasmid construct was made and shown to express a self-regulating bicistronic transcript containing both the reporter and the transactivator. These Tc-controlled plasmids, termed maximum expression and regulated vectors (MERVs), have the potential to target a variety of gene therapy applications.
authors.affiliation: Department of Molecular Biology, Vical Incorporated, San Diego, CA 92121, USA.
authors.forename: X
authors.initials: X
authors.lastname: Liang
authors.forename: J
authors.initials: J
authors.lastname: Hartikka
authors.forename: L
authors.initials: L
authors.lastname: Sukhu
authors.forename: M
authors.initials: M
authors.lastname: Manthorpe
authors.forename: P
authors.initials: P
authors.lastname: Hobart
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics; D015533:Transcriptional Activation
chemical_list: D000900:Anti-Bacterial Agents; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D015500:Chloramphenicol O-Acetyltransferase; D013752:Tetracycline