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An episomal vector for stable tetracycline-regulated gene expression
Journal article   Open access   Peer reviewed

An episomal vector for stable tetracycline-regulated gene expression

M Jost, C Kari and U Rodeck
Nucleic acids research, v 25(15), pp 3131-3134
01 Aug 1997
PMID: 9224615
url
https://doi.org/10.1093/nar/25.15.3131View
Published, Version of Record (VoR) Open

Abstract

Dose-Response Relationship, Drug Gene Expression Regulation - drug effects Transfection Plasmids Humans Tetracycline - pharmacology HeLa Cells Tumor Cells, Cultured Genetic Vectors Genes, Reporter Luciferases - genetics
The recently introduced tetracycline (Tc)-regulatable eukaryotic gene expression system based on the Escherichia coli Tn 10 tetracycline operon has proven to be a powerful tool for controlled expression of a variety of genes in vitro as well as in vivo . Control elements of this expression system are contained in two separate plasmid vectors. The tTA vector encodes a transactivator protein and the tetP vector contains a responsive operator-promoter element (tetP) that controls gene expression depending on tTA binding. Establishment of cell lines expressing a gene of interest under tetP control requires two subsequent rounds of transfection and clonal selection after each transfection. Here we describe a modification of this system in which the tetP element is placed in an episomal EBNA-based plasmid that can be stably maintained in primate but not in rodent cells. Using HeLa and human melanoma cells, we show that upon transient or stable transfection a reporter gene is expressed in a Tc-regulated manner similar to the original system. Thus, this expression system combines the advantages of episomal vectors, such as high efficiency of transfection and time-efficient selection of mass cultures, with tight control of gene expression provided by the Tc-regulatable system.

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Biochemistry & Molecular Biology
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