Tight control of gene expression by a helper-dependent adenovirus vector carrying the rtTA2s-M2 tetracycline transactivator and repressor system

V. Salucci, A. Scarito, L. Aurisicchio, S. Lamartina, G. Nicolaus, S. Giampaoli, O. Gonzalez-Paz, C. Toniatti, H. Bujard, W. Hillen, G. Ciliberto, F. Palombo

Research output: Contribution to journalArticle

Abstract

Control of gene expression for gene therapy application requires the design of a sophisticated system embodying multiple properties. The ideal system should present the following features: (1) low or undetectable gene expression in the absence of inducer; (2) strong expression upon induction; and (3) fast kinetics of induction in the presence of inducers and rapid reversal of induction after its withdrawal. To evaluate these parameters, the features of the latest generation tetracycline-sensitive reverse-transactivator (rtTA2s-M2) alone or in combination with Tet-repressor (tTS-Kid) were explored in the context of helper-dependent adenovirus vector. Various genetic elements were assembled in a series of vectors and the ability to control secreted alkaline phosphatase expression evaluated in vitro in HeLa cells and in vivo by intramuscular injection in both C57/B6 and Balb/C nude mice. The results allow us to draw some general conclusions about the combination of transcription regulators and their relative orientation to the transgene to achieve maximal induction, while minimizing leakiness of expression.

Original languageEnglish
Pages (from-to)1415-1421
Number of pages7
JournalGene Therapy
Volume9
Issue number21
DOIs
Publication statusPublished - Nov 2002

Keywords

  • Gene regulation
  • Helper-dependent adenovirus vectors
  • Muscle
  • Silencer
  • Tetracycline

ASJC Scopus subject areas

  • Genetics

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  • Cite this

    Salucci, V., Scarito, A., Aurisicchio, L., Lamartina, S., Nicolaus, G., Giampaoli, S., Gonzalez-Paz, O., Toniatti, C., Bujard, H., Hillen, W., Ciliberto, G., & Palombo, F. (2002). Tight control of gene expression by a helper-dependent adenovirus vector carrying the rtTA2s-M2 tetracycline transactivator and repressor system. Gene Therapy, 9(21), 1415-1421. https://doi.org/10.1038/sj.gt.3301813