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Turkish Journal of Medical Sciences

DOI

10.3906/sag-1008-1057

Abstract

To describe in detail the use of the T-REx^{TM} System (a tetracycline-regulated mammalian expression system; Invitrogen) for rapid, easy, and effective generation of stable C2C12 cell clones. Tetracycline-inducible stable cell lines are useful tools to study the function of various genes in different types of mammalian cells. However, the generation of stable cell lines is a time-consuming, technically difficult, and expensive process. Materials and methods: Generation of a stable C2C12 cell line was performed by stable transfection of the cells with the pcDNA6/TR plasmid vector, which contains the tetracycline repressor and the blasticidin drug resistance genes. The establishment of the stable cell line and the efficiency of the tetracycline-inducible gene expression system were shown with the transient transfection of reporter plasmid pTO-EGFP and observation of the GFP expression by fluorescence microscopy. Results: We established a stable T-REx^{TM} C2C12 cell line that can be used for muscle research studies by transfecting the gene of interest with a second plasmid. Conclusion: The T-REx^{TM} system is useful in the development of stable cell lines for protein production or regulation of gene expression.

Keywords

C2C12 cell line, stable transfection, inducible expression, tetracycline resistance gene

First Page

821

Last Page

825

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