Dual-regulated expression technology: a new era in the adjustment of heterologous gene expression in mammalian cells

Background On the basis of the compatible streptogramin‐ and tetracycline‐responsive expression systems, a series of dual‐regulated expression systems have been established for use in sophisticated biopharmaceutical manufacturing, advanced gene therapy, and tissue engineering. Methods Dual‐regulated expression concepts enable streptogramin‐ and tetracycline‐responsive control of two different (sets of) transgenes (multiregulated multigene metabolic engineering), dual‐autoregulated expression configurations for one‐step chromosomal integration of two antibiotic‐adjustable expression units, and artificial regulatory cascades for multi‐level regulation of transgenes and optimized integration of molecular interventions into mammalian regulatory networks. Results This report describes the construction and testing of a family of dual‐regulated expression vectors which are compatible with the pTRIDENT vector construction kit, and, in some cases, adapted for retroviral expression technology enabling straightforward transduction of difficult‐to‐transfect cell lines such as primary cells and stem cells. Conclusions Dual‐regulated expression technology will probably become of prime interest for a variety of therapeutic applications, including biopharmaceutical manufacturing, gene therapy, and tissue engineering. Copyright © 2001 John Wiley & Sons, Ltd.