Generation of stable human cell lines with tetracycline-inducible (Tet-on) shRNA or cDNA expression

Journal of Visualized Experiments

Volumn , Issue 73, 2013, Pages

  Gomez Martinez, Marta ^a   Schmitz, Debora ^b   Hergovich, Alexander ^a  

^a UNIVERSITY COLLEGE LONDON   (United Kingdom)

^b FRIEDRICH MIESCHER INSTITUTE FOR BIOMEDICAL RESEARCH   (Switzerland)

Author keywords

Anatomy; Bioengineering; Biomedical Engineering; cDNA; Cell Biology; Cell culture; Cellular Biology; Clones; DNA; Expression; Genes; Genetics; Issue 73; Mammals; Medicine; Molecular Biology; Physiology; Promoter; Proteins; shRNA; Stable manipulation of cell lines; Tetracycline; Tetracycline regulated expression; Tissue culture; Vectors

Indexed keywords

COMPLEMENTARY DNA; SMALL INTERFERING RNA; TETRACYCLINE;

ARTICLE; BIOSYNTHESIS; CELL LINE; GENE EXPRESSION REGULATION; GENE SILENCING; GENETIC PROCEDURES; GENETICS; HUMAN;

CELL LINE; DNA, COMPLEMENTARY; GENE EXPRESSION REGULATION; GENE KNOCKDOWN TECHNIQUES; GENETIC TECHNIQUES; HUMANS; RNA, SMALL INTERFERING; TETRACYCLINE;

EID: 84875336693     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/50171     Document Type: Article

References (17)

1. Postle, K., Nguyen, T.T., & Bertrand, K.P. Nucleotide sequence of the repressor gene of the TN10 tetracycline resistance determinant. Nucleic Acids Res. 12, 4849-4863 (1984).
2. Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U.S.A. 89, 5547-5551 (1992).
3. Gossen, M., et al. Transcriptional activation by tetracyclines in mammalian cells. Science. 268, 1766-1769 (1995).
4. Elbashir, S.M., et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411, 494-498, doi:10.1038/35078107 (2001).
5. Brummelkamp, T.R., Bernards, R., & Agami, R. A system for stable expression of short interfering RNAs in mammalian cells. Science. 296, 550-553, doi:10.1126/science.1068999 (2002).
6. McManus, M.T., Petersen, C.P., Haines, B.B., Chen, J., & Sharp, P.A. Gene silencing using micro-RNA designed hairpins. RNA. 8, 842-850 (2002).
7. Miyagishi, M. & Taira, K. U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells. Nat. Biotechnol. 20, 497-500, doi:10.1038/nbt0502-497 (2002).
8. Paddison, P.J., Caudy, A.A., Bernstein, E., Hannon, G.J., & Conklin, D.S. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev. 16, 948-958, doi:10.1101/gad.981002 (2002).
9. Paul, C.P., Good, P.D., Winer, I., & Engelke, D.R. Effective expression of small interfering RNA in human cells. Nat. Biotechnol. 20, 505-508, doi:10.1038/nbt0502-505 (2002).
10. Sui, G., et al. A DNA vector-based RNAi technology to suppress gene expression in mammalian cells. Proc. Natl. Acad. Sci. U.S.A. 99, 5515-5520, doi:10.1073/pnas.082117599 (2002).
11. Yu, J.Y., DeRuiter, S.L., & Turner, D.L. RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells. Proc. Natl. Acad. Sci. U.S.A. 99, 6047-6052, doi:10.1073/pnas.092143499 (2002).
12. van de Wetering, M., et al. Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. EMBO Rep. 4, 609-615, doi:10.1038/sj.embor.embor865 (2003).
13. Hergovich, A., et al. The MST1 and hMOB1 tumor suppressors control human centrosome duplication by regulating NDR kinase phosphorylation. Curr. Biol. 19, 1692-1702, doi:10.1016/j.cub.2009.09.020 (2009).
14. Hergovich, A., Lamla, S., Nigg, E.A., & Hemmings, B.A. Centrosome-associated NDR kinase regulates centrosome duplication. Mol. Cell. 25, 625-634, doi:10.1016/j.molcel.2007.01.020 (2007).
15. Kohler, R.S., Schmitz, D., Cornils, H., Hemmings, B.A., & Hergovich, A. Differential NDR/LATS interactions with the human MOB family reveal a negative role for human MOB2 in the regulation of human NDR kinases. Mol. Cell Biol. 30, 4507-4520, doi:10.1128/MCB.00150-10 (2010).
16. Vichalkovski, A., et al. NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr. Biol. 18, 1889-1895, doi:10.1016/j.cub.2008.10.060 (2008).
17. Pear, W.S., et al. Efficient and rapid induction of a chronic myelogenous leukemia-like myeloproliferative disease in mice receiving P210 bcr/abl-transduced bone marrow. Blood. 92, 3780-3792 (1998).