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Streptogramin-based gene regulation systems for mammalian cells
M Fussenegger1, R P Morris, C Fux
1Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland. fussen@biotech.biol.ethz.ch
Nature Biotechnology
|November 4, 2000
Summary
Researchers developed new gene expression systems (PipON and PipOFF) using pristinamycin-induced protein (Pip) that are controlled by streptogramin antibiotics. These systems offer improved performance over existing methods and allow for independent control of multiple genes.
Area of Science:
- Molecular Biology
- Gene Regulation
- Biotechnology
Background:
- Regulated gene expression is crucial for controlling cellular functions and developing novel therapeutics.
- Existing systems like tetracycline-repressible systems have limitations in background expression and induction ratios.
Purpose of the Study:
- To develop novel inducible and repressible gene expression systems in mammalian cells.
- To utilize the pristinamycin-induced protein (Pip) repressor for streptogramin-antibiotic-controlled gene expression.
- To compare the efficacy of the new Pip-based systems with existing tetracycline-repressible systems.
Main Methods:
- Development of PipON (inducible) and PipOFF (repressible) gene expression systems.
- Utilizing the Pip repressor from Streptomyces coelicolor.
- Testing gene expression of erythropoietin (EPO), secreted alkaline phosphatase (SEAP), and green fluorescent protein (GFP) in various mammalian cell lines (BHK, CHO, HeLa, mouse myoblasts).
- Comparative analysis with tetracycline-repressible (TetOFF) systems in CHO cells.
Main Results:
- Streptogramin antibiotics (pristinamycin, virginiamycin, Synercid) effectively regulated gene expression in PipON and PipOFF systems.
- Pip-based systems demonstrated versatility across multiple cell types and for different reporter genes.
- The PipOFF system exhibited lower background expression and higher induction ratios compared to TetOFF systems.
- Functional compatibility between streptogramin-based and TetOFF systems was confirmed, enabling dual-gene control.
Conclusions:
- The PipON and PipOFF systems provide a novel, antibiotic-inducible/repressible method for gene expression in mammalian cells.
- These systems offer advantages over existing technologies, including improved performance and dual-control capabilities.
- Streptogramin-based gene regulation presents a promising tool for research and therapeutic applications.