title: Development and validation of a robust and versatile one-plasmid regulated gene expression system.
abstract: We have developed a one-plasmid regulated gene expression system, pBRES, based on a mifepristone (MFP)-inducible two-plasmid system. The various expression elements of the pBRES system (promoters, 5' and 3' untranslated regions (UTRs), introns, target gene, and polyA sequences) are bounded by restriction enzyme sites so that each module can be conveniently replaced by alternate DNA elements in order to tailor the system for particular tissues, organs, or conditions. There are four possible orientations of the two expression units relative to each other, and insertion of a variety of expression elements and target genes into the four different orientations revealed orientation- and gene-dependent effects on induced and uninduced levels of gene expression. Induced target gene expression from the pBRES system was shown to be comparable to the two-plasmid system and higher than the expression from the cytomegalovirus (CMV) promoter in vivo, while maintaining low uninduced levels of expression. Finally, a pBRES expression cassette was transferred to an adeno-associated virus (AAV) vector and shown to be capable of regulated gene expression in vivo for nearly 1 year.
authors.affiliation: Department of Gene Technologies, Berlex Biosciences, Richmond, California, USA. Paul.Szymanski@bayer.com
authors.forename: Paul
authors.initials: P
authors.lastname: Szymanski
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Kretschmer
authors.forename: Maxine
authors.initials: M
authors.lastname: Bauzon
authors.forename: Fang
authors.initials: F
authors.lastname: Jin
authors.forename: Hu Sheng
authors.initials: HS
authors.lastname: Qian
authors.forename: Gabor M
authors.initials: GM
authors.lastname: Rubanyi
authors.forename: Richard N
authors.initials: RN
authors.lastname: Harkins
authors.forename: Terry W
authors.initials: TW
authors.lastname: Hermiston
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016899:Interferon-beta / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics*; D019076:Transgenes / Q000235:genetics
chemical_list: D016899:Interferon-beta