Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism.

  • Authors:
    • H Veelken
    • B Leutgeb
    • P Kulmburg
    • H H Fiebig
    • A Mackensen
    • A Lindemann
  • View Affiliations

  • Published online on: October 1, 1998     https://doi.org/10.3892/ijmm.2.4.423
  • Pages: 423-431
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Abstract

Success of gene replacement therapy depends on long-term, high level expression of the transgene. Gene therapy vectors incorporating a promoter of a constitutively active eukaryotic gene may allow long-term expression in vivo, but the expression level may be insufficient for therapeutic effects. To enhance transcription from eukaryotic promoters, a strategy with dicistronic vectors encoding the therapeutic gene of interest together with a transcription factor that binds and activates the promoter was tested. Expression vectors for the chimeric tet repressor/VP16 transcription factor (tTA) driven by the human beta-actin promoter were constructed, and tandem tet operators were inserted within the promoter. This arrangement significantly enhanced expression of G-CSF in fibroblasts to higher levels than the immediate/early CMV promoter. Stably transfected fibroblast clones produced up to 2.4 microg G-CSF per 10(6) cells x 24 h. After injection of genetically engineered cells into SCID mice, the enhanced beta-actin promoter construct resulted in marked leukocytosis, whereas the unmodified promoter had only a marginal therapeutic effect. Transcription factor-enhanced, feed-back-activated human promoters may thus achieve higher expression levels than viral control elements, and may be advantageous for gene therapy due to high constitutive activity in vivo.

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Oct 1998
Volume 2 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Veelken H, Leutgeb B, Kulmburg P, Fiebig H, Mackensen A and Lindemann A: Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism.. Int J Mol Med 2: 423-431, 1998
APA
Veelken, H., Leutgeb, B., Kulmburg, P., Fiebig, H., Mackensen, A., & Lindemann, A. (1998). Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism.. International Journal of Molecular Medicine, 2, 423-431. https://doi.org/10.3892/ijmm.2.4.423
MLA
Veelken, H., Leutgeb, B., Kulmburg, P., Fiebig, H., Mackensen, A., Lindemann, A."Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism.". International Journal of Molecular Medicine 2.4 (1998): 423-431.
Chicago
Veelken, H., Leutgeb, B., Kulmburg, P., Fiebig, H., Mackensen, A., Lindemann, A."Enhancement of a constitutively active promoter for gene therapy by a positive feed-back transcriptional activator mechanism.". International Journal of Molecular Medicine 2, no. 4 (1998): 423-431. https://doi.org/10.3892/ijmm.2.4.423