Gas-inducible transgene expression in mammalian cells and mice.
We describe the design and detailed characterization of a gas-inducible transgene control system functional in different mammalian cells, mice and prototype biopharmaceutical manufacturing. The acetaldehyde-inducible AlcR-P(alcA) transactivator-promoter interaction of the Aspergillus nidulans ethanol-catabolizing regulon was engineered for gas-adjustable transgene expression in mammalian cells. Fungal AlcR retained its transactivation characteristics in a variety of mammalian cell lines and reversibly adjusted transgene transcription from chimeric mammalian promoters (P(AIR)) containing P(alcA)-derived operators in a gaseous acetaldehyde-dependent manner. Mice implanted with microencapsulated cells engineered for acetaldehyde-inducible regulation (AIR) of the human glycoprotein secreted placental alkaline phosphatase showed adjustable serum phosphatase levels after exposure to different gaseous acetaldehyde concentrations. AIR-controlled interferon-beta production in transgenic CHO-K1-derived serum-free suspension cultures could be modulated by fine-tuning inflow and outflow of acetaldehyde-containing gas during standard bioreactor operation. AIR technology could serve as a tool for therapeutic transgene dosing as well as biopharmaceutical manufacturing.
Weber
Wilfried
W
Institute for Chemical and Bio-Engineering, Swiss Federal Institute of Technology, ETH Hoenggerberg HCI F115, CH-8093 Zurich, Switzerland.
Rimann
Markus
M
Spielmann
Manuela
M
Keller
Bettina
B
Daoud-El Baba
Marie
M
Aubel
Dominique
D
Weber
Cornelia C
CC
Fussenegger
Martin
M
D000079:Acetaldehyde / Q000494:pharmacology*; D000469:Alkaline Phosphatase; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D005260:Female; D058851:GPI-Linked Proteins; D005740:Gases / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D024861:Genetic Enhancement / Q000379:methods; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics*; D051379:Mice; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D019076:Transgenes / Q000187:drug effects*
D058851:GPI-Linked Proteins; D005740:Gases; D007527:Isoenzymes; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental; D000079:Acetaldehyde