An Improved Vector System for Homogeneous and Stable Gene Regulation
| Autor: | Adolfo Rivero-Müller, Kamil Filipek, Patrycja Rulak, Marek Tchórzewski, Barbara Michalec-Wawiórka, Jakub Czapinski, Arkadiusz Czerwonka |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Ribosomal Protein L10
QH301-705.5 Transgene Genetic Vectors Green Fluorescent Proteins Computational biology Biology Catalysis Article Inorganic Chemistry Transactivation Mice Ribosomal protein Gene expression Animals Humans expression systems Transgenes Physical and Theoretical Chemistry Biology (General) Molecular Biology Gene QD1-999 Spectroscopy Regulation of gene expression Tet-On uL10 (P0) ribosomal proteins Organic Chemistry General Medicine biochemical phenomena metabolism and nutrition Tetracycline Computer Science Applications Anti-Bacterial Agents Chemistry HEK293 Cells Expression (architecture) Gene Expression Regulation Genetic Techniques Homogeneous Doxycycline NIH 3T3 Cells Trans-Activators gene regulation HeLa Cells |
| Zdroj: | International Journal of Molecular Sciences, Vol 22, Iss 5206, p 5206 (2021) International Journal of Molecular Sciences Volume 22 Issue 10 |
| ISSN: | 1661-6596 1422-0067 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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