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  • Title: An Improved Vector System for Homogeneous and Stable Gene Regulation.
    Author: Michalec-Wawiórka B, Czapiński J, Filipek K, Rulak P, Czerwonka A, Tchórzewski M, Rivero-Müller A.
    Journal: Int J Mol Sci; 2021 May 14; 22(10):. PubMed ID: 34069024.
    Abstract:
    Precise analysis of the genetic expression and functioning of proteins requires experimental approaches that, among others, enable tight control of gene expression at the transcriptional level. Doxycycline-induced Tet-On/Tet-Off expression systems provide such an opportunity, and are frequently used to regulate the activity of genes in eukaryotic cells. Since its development, the Tet-system has evolved tight gene control in mammalian cells; however, some challenges are still unaddressed. In the current set up, the establishment of the standard Tet-based system in target cells is time-consuming and laborious and has been shown to be inefficient, especially in a long-term perspective. In this work, we present an optimized inducible expression system, which enables rapid generation of doxycycline-responsive cells according to a one- or two-step protocol. The reported modifications of the Tet-On system expand the toolbox for regulated mammalian gene expression and provide high, stable, and homogenous expression of the Tet-On3G transactivator, which is of fundamental importance in the regulation of transgenes.
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