Nature Biotechnology
20, 901 - 907 (2002)
Published online: 19 August 2002; | doi:10.1038/nbt731
Macrolide-based transgene control in mammalian cells and miceWilfried Weber1, Cornelia Fux1, Marie Daoud-El Baba2, Bettina Keller1, Cornelia C. Weber3, Beat P. Kramer1, Christoph Heinzen4, Dominique Aubel2, James E. Bailey1, 5
& Martin Fussenegger11
Institute of Biotechnology, Swiss Federal
Institute of Technology, ETH Zurich, CH-8093
Zurich, Switzerland. 2
Institut Universitaire de Technologie, IUTA,
Département Génie Biologique, 43 Boulevard du 11
Novembre 1918, F-69200 Villeurbanne Cedex,
France. 3
Institute for Biomedical Engineering, Swiss
Federal Institute of Technology, ETH Zurich, CH-8044
Zurich, Switzerland. 4
Inotech Encapsulation AG,
Kirchstrasse 1, CH-5605 Dottikon,
Switzerland. 5
Deceased.
Correspondence should be addressed to Martin Fussenegger fussenegger@biotech.biol.ethz.chHeterologous mammalian gene regulation systems for adjustable
expression of multiple transgenes are necessary for advanced human gene therapy
and tissue engineering, and for sophisticated in vivo gene-function
analyses, drug discovery, and biopharmaceutical manufacturing. The
antibiotic-dependent interaction between the repressor (E) and operator (ETR)
derived from an Escherichia coli erythromycin-resistance regulon was
used to design repressible (EOFF) and inducible (EON)
mammalian gene regulation systems (E.REX) responsive to clinically licensed
macrolide antibiotics (erythromycin, clarithromycin, and roxithromycin). The
EOFF system consists of a chimeric erythromycin-dependent
transactivator (ET), constructed by fusing the prokaryotic repressor E to a
eukaryotic transactivation domain that binds and activates transcription from
ETR-containing synthetic eukaryotic promoters (PETR). Addition of
macrolide antibiotic results in repression of transgene expression. The
EON system is based on E binding to artificial ETR-derived operators
cloned adjacent to constitutive promoters, resulting in repression of transgene
expression. In the presence of macrolides, gene expression is induced. Control
of transgene expression in primary cells, cell lines, and microencapsulated
human cells transplanted into mice was demonstrated using the E.REX
(EOFF and EON) systems. The macrolide-responsive E.REX
technology was functionally compatible with the streptogramin
(PIP)−regulated and tetracycline (TET)−regulated expression
systems, and therefore may be combined for multiregulated multigene therapeutic
interventions in mammalian cells and tissues.
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