A novel vector platform for vitamin H-inducible transgene expression in mammalian cells.

A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Research Abstract Details 

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A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Abstract Text:

Wilfried Weber,William Bacchus,Franziska Gruber,Mattia Hamberger,Martin Fussenegger,

Inducible transgene control systems have been instrumental to gene therapy, biopharmaceutical manufacturing, drug discovery, synthetic biology and functional genomic research. The most widely used heterologous gene regulation systems are responsive to antibiotics of the tetracycline, streptogramin and macrolide classes. Although these antibiotics are clinically licensed, concerns about the emergence of resistant bacteria, side-effects in animal studies, and economic considerations associated with clearance of antibiotics in biopharmaceutical manufacturing, have limited the use of heterologous transgene control modalities to basic research activities. We have therefore designed a strategy to convert antibiotic-responsive transcription factors into gene regulation systems responsive to non-toxic biotin, also known as vitamin H. Constitutive ligation of biotin to the Avitag-containing VP16 transactivation domain by the Escherichia coli biotin ligase BirA enables heterodimerization with tetracycline- (TetR), streptogramin- (Pip), and macrolide- (E) dependent repressors fused to streptavidin, which creates synthetic transactivators able to activate specific promoters (P(hCMV-1), P(PIR), P(ETR)). We have demonstrated (i) that exogenous biotin (40nM) can induce heterologous transgene expression in a biotin- (serum-) free culture environment (biotin-dependent heterodimerization of transactivator); (ii) that excess biotin (above 200microM) gradually represses transgene expression in a biotin- (serum-) containing environment (saturation of streptavidin by excess biotin prevents heterodimerization of the transactivator); and (iii) that avidin can sequestrate endogenous biotin in serum-containing cultures and so repress transgene expression in a dose-dependent manner. In addition, by engineering all off the components required for biotin-controlled transgene expression (Avitag-VP16, repressor-streptavidin, BirA) into a tricistronic (lenti)vector configuration, it was possible to transfect (transduce) a variety of mammalian cell lines and primary cells and enable biotin-controlled transgene expression in a simple and straightforward manner. The conversion of generic antibiotic-responsive transcription control modalities into systems adjustable by non-toxic vitamin H may foster novel advances in reprogramming of mammalian cells and production of difficult-to-produce protein pharmaceuticals.

A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Publishing Authors By Initials

W Weber,W Bacchus,F Gruber,M Hamberger,M Fussenegger,

For similar viruses: virion research abstracts see: viruses: virion research

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A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biotechnology

VOLUME: 131

Page Numbers: 150-8

Journal Abbreviation: J. Biotechnol.

ISSN: 0168-1656

DAY: 23

MONTH: 06

YEAR: 2007

A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Information

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LANGUAGE: eng

NlmUniqueID: 8411927

A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Keywords Mesh Terms:

KEYWORDS: Virion

MESH TERMS: genetics

Chemical & Substance for Abstract: A novel vector platform for vitamin H-inducible transgene expression in mammalian cells. Information

Substance Name: Biotin

Registry Number: 58-85-5

Grant and Affiliation Information for A novel vector platform for vitamin H-inducible transgene expression in mammalian cells.

AFFILIATION: Institute for Chemical and Bioengineering, ETH Zurich, HCI F115, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland.

Country: Netherlands

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MEDLINETA: J Biotechnol

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