Versatile macrolide-responsive mammalian expression vectors for multiregulated multigene metabolic engineering

• Published in: Biotechnology and bioengineering 2002-12, Vol.80 (6), p.691-705
• Main Authors: Weber, Wilfried, Marty, René R., Keller, Bettina, Rimann, Markus, Kramer, Beat P., Fussenegger, Martin
• Format: Article
• Language: English
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Biological and medical sciences; Biotechnology; CHO Cells - drug effects; CHO Cells - physiology; Cloning, Molecular - methods; Cricetinae; dual-regulated expression; E.REX; erythromycin; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation - drug effects; gene regulation system; gene therapy; Genetic engineering; Genetic Engineering - methods; Genetic technics; Genetic Therapy - methods; Genetic Vectors; macrolide antibiotics; Macrolides - pharmacology; Methods. Procedures. Technologies; Multigene Family; pDuoRex; PIP system; pRetroTriRex; pTRIDENT; pTriRex; pTwin; streptogramin antibiotics; Streptogramins - pharmacology; TET system; Tetracycline - pharmacology; tetracycline antibiotics; tissue engineering; Tissue Engineering - methods; Transcriptional Activation - drug effects; Transgenes - drug effects; Transgenes - genetics; triple-regulated expression; Vectors (cloning, transfer, expression). Insertion sequences and transposons
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.10461

The novel macrolide‐inducible and ‐repressible mammalian gene regulation systems (E.REX) have been cloned into a variety of sophisticated expression configurations including (1) multi‐purpose expression vectors, (2) pTRIDENT‐based artificial operons, (3) dual‐regulated expression strategies for independent control of two different transgenes, (4) autoregulated vectors for one‐step installation of adjustable multigene expression, and (5) oncoretroviral and lentiviral plasmids for transduction of macrolide‐, streptogramin‐ and tetracycline‐dependent transactivators and production of cell lines supporting independent control of three different transgenes. This vector portfolio represents a construction kit‐like toolbox for efficient installation of adjustable gene expression responsive to clinically licensed antibiotics and enables the design of multiregulated multigene metabolic engineering strategies required for biopharmaceutical manufacturing, gene therapy, and tissue engineering. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 691–705, 2002.