Generation of cells expressing improved doxycycline-regulated reverse transcriptional transactivator rtTA2S-M2

Nature Protocols

Volumn 1, Issue 2, 2006, Pages 803-811

  Welman, Arkadiusz ^a   Barraclough, Jane ^a   Dive, Caroline ^a  

^a UNIVERSITY OF MANCHESTER   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

DOXYCYCLINE; TRANSACTIVATOR PROTEIN;

ARTICLE; CELL LINE; DRUG EFFECT; ELECTROPORATION; FLOW CYTOMETRY; GENE EXPRESSION REGULATION; GENETIC ENGINEERING; GENETIC TRANSFECTION; GENETICS; HUMAN; METABOLISM; METHODOLOGY; PLASMID;

CELL LINE; DOXYCYCLINE; ELECTROPORATION; FLOW CYTOMETRY; GENE EXPRESSION REGULATION; GENETIC ENGINEERING; HUMANS; PLASMIDS; TRANS-ACTIVATORS; TRANSFECTION;

EID: 33750701914     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2006.117     Document Type: Article

References (18)

1. Fussenegger, M. The impact of mammalian gene regulation concepts on functional genomic research, metabolic engineering, and advanced gene therapies. Biotechnol. Prog. 17, 1-51 (2001).
2. Kaufmann, H. & Fussenegger, M. Metabolic engineering of mammalian cells for higher protein yieldin. Gene Transfer and Expression in Mammalian Cells (ed. Makrides, S.C.) 457-569 (Elsevier Science, Amsterdam, 2003).
3. Toniatti, C., Bujard, H., Cortese, R. & Ciliberto, G. Gene therapy progress and prospects: Transcription regulatory systems. Gene Ther. 21, 649-657 (2004).
4. Berens, C. & Hillen, W. Gene regulation by tetracyclines. Constraints of resistance regulation in bacteria shape TetR for application in eukaryotes. Eur. J. Biochem. 270, 3109-3121 (2003).
5. Gossen, M. et al. Transcriptional activation by tetracyclines in mammalian cells. Science 268, 1766-1769 (1995).
6. Corbel, S.Y. & Rossi, F. M. Latest developments and in vivo use of the Tet system: ex vivo and in vivo delivery of tetracycline-regulated genes. Curr. Opin. Biotechnol. 13, 448-452 (2002).
7. Zhu, Z., Zheng, T., Lee, C.G., Homer, R.J. & Elias, J.A. Tetracycline-controlled transcriptional regulation systems: Advances and application in transgenic ardmat modeling. Semin. Cell. Dev. Biol. 13, 121-128 (2002).
8. Baron, U. & Bujard, H. Tet repressor-based system for regulated gene expression in eukaryotic cells: Principles and advances. Methods Enzymol. 327, 401-421 (2000).
9. Urlinger, S. et al. Exploring the sequence space for tetracycline-dependent transcriptional activators: Novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. USA 97 7963-7968 (2000).
10. Hofmann, A., Nolan, G.P. & Blau, H.M. Rapid retrovirat delivery of tetracycline-inducible genes in a single autoregulatory cassette. Proc. Natl. Acad. Sci. USA 93, 5185-5190 (1996).
11. Chtarto, A. et al. Tetracycline-inducibie transgene expression mediated by a single AAV vector. Gene Ther. 10, 84-94 (2003).
12. Szulc, J., Wiznerowicz, M., Sauvain, M.O., Trono, D. & Aebischer, P. A versatile tool for conditional gene expression and knockdown. Nat. Methods 3, 109-116 (2006).
13. Gopalkrishnan, R.V., Christiansen, K.A., Goldstein, N.I., DePinho, R.A. & Fisher, P.B. Use of the human EF-1α promoter for expression can significantly increase success in establishing stable cell lines with consistent expression: A study using the tetracycline-inducible system in human cancer cells. Nucleic Acids Res. 27, 4775-4782 (1999).
14. Munoz, I., Gomez, A., Zanuy, S. & Carrillo, M. A one-step approach to obtain cell clones expressing tetracycline-responsive transactivators. Anal. Biochem. 331, 153-160 (2004).
15. Whitelaw, E. et al. Epigenetic effects on transgene expression. Methods Mol. Biol. 158, 351-368 (2001).
16. Welman, A. et al. Construction and characterization of multiple human colon cancer cell lines for inducibly regulated gene expression. J. Cell. Biochem. 94, 1148-1162 (2005).
17. Qu, Z. et al. Homogeneity and long-term stability of tetracycline-regulated gene expression with low basal activity by using the rtTA2S-M2 ransactivator and insulator-flanked reporter vectors. Gene 327, 61-73 (2004).
18. Mizuguchi, H., Xu, Z.L., Sakurai, F., Mayumi, T. & Hayakawa, T. Tight positive regulation of transgene expression by a single adenovirus vector containing the rtTA and tTS expression cassettes in separate genome regions. Hum. Gene Ther. 14, 1265-1277 (2003).