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 (multi-regulated 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.
Moser
S
S
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich.
Rimann
M
M
Fux
C
C
Schlatter
S
S
Bailey
J E
JE
Fussenegger
M
M
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D005246:Feedback; D005786:Gene Expression Regulation / Q000187:drug effects; D005821:Genetic Techniques; D005822:Genetic Vectors; D008322:Mammals; D025762:Pristinamycin / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics; D025361:Streptogramins / Q000494:pharmacology; D013754:Tetracyclines / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics; D015533:Transcriptional Activation; D014162:Transfection; D019076:Transgenes
D025762:Pristinamycin; D025361:Streptogramins; D013754:Tetracyclines; D015534:Trans-Activators