Dual-regulated expression technology: a new era in the adjustment of heterologous gene expression in mammalian cells

J Gene Med. 2001 Nov-Dec;3(6):529-49. doi: 10.1002/jgm.219.

Abstract

Background: On the basis of the compatible streptogramin- and tetracycline-responsive expression systems, a series of dual-regulated expression systems have been established for use in sophisticated biopharmaceutical manufacturing, advanced gene therapy, and tissue engineering.

Methods: Dual-regulated expression concepts enable streptogramin- and tetracycline-responsive control of two different (sets of) transgenes (multi-regulated multigene metabolic engineering), dual-autoregulated expression configurations for one-step chromosomal integration of two antibiotic-adjustable expression units, and artificial regulatory cascades for multi-level regulation of transgenes and optimized integration of molecular interventions into mammalian regulatory networks.

Results: This report describes the construction and testing of a family of dual-regulated expression vectors which are compatible with the pTRIDENT vector construction kit, and, in some cases, adapted for retroviral expression technology enabling straightforward transduction of difficult-to-transfect cell lines such as primary cells and stem cells.

Conclusions: Dual-regulated expression technology will probably become of prime interest for a variety of therapeutic applications, including biopharmaceutical manufacturing, gene therapy, and tissue engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotechnology / methods*
  • CHO Cells
  • Cells, Cultured
  • Cloning, Molecular
  • Cricetinae
  • Feedback
  • Gene Expression Regulation* / drug effects
  • Genetic Techniques*
  • Genetic Vectors
  • Mammals
  • Pristinamycin / pharmacology
  • Promoter Regions, Genetic
  • Retroviridae / genetics
  • Streptogramins / pharmacology
  • Tetracyclines / pharmacology
  • Trans-Activators / genetics
  • Transcriptional Activation
  • Transfection
  • Transgenes

Substances

  • Pristinamycin
  • Streptogramins
  • Tetracyclines
  • Trans-Activators