The DOI matches its Crossref record; 39 of the 39 references it cites were found in a registry or an index.
Tetracycline-Reversible Silencing of Eukaryotic Promoters
Open accessAbstract
A tetracycline-controlled transrepressor protein has been engineered to silence transcriptional activities of eukaryotic promoters that are stably integrated into the chromatin of human cells. By fusing the KRAB domain of human Kox1 to the Tet repressor derived from Tn10 of Escherichia coli, a tetracycline-controlled hybrid protein (TetR-KRAB) was generated and constitutively expressed in HeLa cells. The TetR-KRAB protein binds to tet operator (tetO) sequences in the absence but not in the presence of tetracycline. When TetR-KRAB bound to tetO sequences upstream of the immediate-early promoter-enhancer of human cytomegalovirus (CMV), the expression of a CMV-driven luciferase reporter construct (ptetO7-CMV-L) was repressed in transient transfection experiments. This silencing was found to operate on different promoters and from tetO sequences placed more than 3 kb from the transcriptional start site. We constructed a stable, doubly transfected cell line (TIS-10) carrying a chromosomally integrated ptetO7-CMV-L reporter construct and expressing the TetR-KRAB protein. Upon addition of tetracycline, luciferase expression was induced more than 50-fold above the baseline level, with half-maximal induction by 2 days. Furthermore, a protein of around 110 kDa was found to coimmunoprecipitate with the TetR-KRAB fusion protein. This protein might play a role as an adaptor protein mediating the silencing exerted by the TetR-KRAB protein. The TetR-KRAB silencing system should be useful as a genetic switch for regulating the expression of chromosomally integrated heterologous and endogenous genes present in mammalian genomes.
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Cites
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“In contrast to a previously published system for highly efficient regulation of gene expression using a tTA-responsive promoter (15, 16), the tetracycline-reversible silencing of complex promoters by the TetR-KRAB protein offers the unique possibility of reversibly down-regulating the expression of cellular genes on t…”
BackgroundMethodInfluentialFirefly luciferase gene: structure and expression in mammalian cells.
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Cited by
CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes
Gilbert, L. A., Larson, M. H., Morsut, L., Liu, Z., Brar, G. A., Torres, S. E., Stern-Ginossar, N., Brandman, O., Whitehead, E. H., Doudna, J. A., et al. (2013) · Cell · doi:10.1016/j.cell.2013.06.044 · Cited by 3,696
“the TALE proteins, have been fused to transcription activators and repressors to modulate gene expression (Cong et al., 2012; Deuschle et al., 1995; Gossen andBujard, 1992; Hathaway et al., 2012; Maeder et al., 2013; Margolin et al., 1994; PerezPinera et al., 2013; Sadowski et al., 1988; Zhang et al., 2000).”
Background5-HT1A autoreceptor levels determine vulnerability to stress and response to antidepressants
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Baron, U., & Bujard, H. (2000) · Methods in Enzymology · doi:10.1016/s0076-6879(00)27292-3 · Cited by 335
Metadata
- Authors
- Deuschle, U., Meyer, W. K.-H., & Thiesen, H.-J.
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 1995-04-01
- Volume
- 15
- Issue
- 4
- Pages
- 1907-1914
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- OpenAlex
- W2142529970
- Cited by
- 265
- References
- 39
- FWCI
- 4.86
- Type
- journal article
- Language
- en
- Open access
- green
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- Deuschle, U., Meyer, W. K.-H., & Thiesen, H.-J. (1995). Tetracycline-Reversible Silencing of Eukaryotic Promoters. Molecular and Cellular Biology. https://doi.org/10.1128/mcb.15.4.1907
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