New-generation multicistronic expression platform
pTRIDENT vectors containing size-optimized IRES elements enable homing endonuclease-based cistron swapping into lentiviral expression vectors
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2004-04
Publication Type
Journal Article
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Abstract
Capitalizing on a proven multicistronic expression vector platform we have designed novel pTRIDENT vectors which (1) enable coordinated expression of three desired transgenes, (2) are size‐optimized, (3) take advantage of small highly efficient internal ribosome entry sites of the GTX or Rbm3 type, (4) harbor various sites specific for homing endonucleases facilitating promoter/multicistronic expression unit/polyadenylation site swapping as well as (5) straightforward integration into human HIV‐l‐based lentiviral expression vectors tailored to contain compatible homing endonucleases. Multicistronic expression profiles of novel pTRIDENT vectors engineered for different tricistronic expression configurations encoding human low‐molecular‐weight urokinase‐type plasminogen activator (u‐PALMW) or Bacillus stearothermophilus‐derived α‐amylase (SAMY), human vascular endothelial growth factor (hVEGF), and human placental secreted alkaline phosphatase (SEAP) have been quantified in Chinese hamster ovary cells (CHO‐K1), mouse fibroblasts (NIH/3T3), and/or human fibrosarcoma (HT‐1080) cells. In addition, a pTRIDENT‐derived SAMY‐VEGF‐SEAP expression cassette transferred into a compatible lentiviral expression vector enabled simultaneous high‐level transgene expression following transduction of transgenic lentiviral particles into primary human chondrocytes. © 2004 Wiley Periodicals, Inc.
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published
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86 (2)
Pages / Article No.
174 - 187
Publisher
Wiley
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Subject
Multicistronic; Metabolic engineering; Multi-gene metabolic engineering; pTRIDENT; Internal ribosome entry site; Rbm3; GTX; homing endonucleases; Lentivirus; SEAP; VEGF; u-PA(LMW); SAMY; CHO-K1; HT-1080; Chondrocytes
Organisational unit
03663 - Aebersold, Rudolf (emeritus) / Aebersold, Rudolf (emeritus)
Notes
Article first published online 19 February 2004, Manuscript accepted 6 November 2003, Manuscript revised 17 June 2003.