Linked Life Data

17669538

Source:http://linkedlifedata.com/resource/pubmed/id/17669538

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2007-8-7
pubmed:abstractText
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.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0168-1656
pubmed:author
pubmed:issnType
Print
pubmed:day
31
pubmed:volume
131
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
150-8
pubmed:meshHeading
pubmed:year
2007
pubmed:articleTitle
A novel vector platform for vitamin H-inducible transgene expression in mammalian cells.
pubmed:affiliation
Institute for Chemical and Bioengineering, ETH Zurich, HCI F115, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Evaluation Studies