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Titolo:
Tetracycline-regulated gene expression mediated by a novel chimeric repressor that recruits histone deacetylases in mammalian cells
Autore:
Jiang, W; Zhou, L; Breyer, B; Feng, T; Cheng, HW; Haydon, R; Ishikawa, A; He, TC;
Indirizzi:
Univ Chicago, Med Ctr, Oncol Mol Lab, Dept Surg, Chicago, IL 60637 USA Univ Chicago Chicago IL USA 60637 l Lab, Dept Surg, Chicago, IL 60637 USA Chongqing Univ Med Sci, Dept Biochem & Mol Biol, Chongqing 400046, PeoplesR China Chongqing Univ Med Sci Chongqing Peoples R China 400046 , PeoplesR China Yamagata Univ, Sch Med, Dept Orthopaed Surg, Yamagata 9909585, Japan Yamagata Univ Yamagata Japan 9909585 opaed Surg, Yamagata 9909585, Japan
Titolo Testata:
JOURNAL OF BIOLOGICAL CHEMISTRY
fascicolo: 48, volume: 276, anno: 2001,
pagine: 45168 - 45174
SICI:
0021-9258(20011130)276:48<45168:TGEMBA>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
TN10 TET REPRESSOR; MAMMARY-TUMOR VIRUS; INDUCIBLE GENE; TRANSGENIC MICE; TRANSCRIPTIONAL REGULATION; IN-VIVO; TRANSACTIVATOR SYSTEM; HUMAN CYTOMEGALOVIRUS; CULTURED-CELLS; PROMOTER;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
66
Recensione:
Indirizzi per estratti:
Indirizzo: He, TC Univ Chicago, Med Ctr, Oncol Mol Lab, Dept Surg, 5841 S Maryland Ave,MC 3079,Rm J-611, Chicago, IL 60637 USA Univ Chicago 5841 S Maryland Ave,MC 3079,Rm J-611 Chicago IL USA 60637
Citazione:
W. Jiang et al., "Tetracycline-regulated gene expression mediated by a novel chimeric repressor that recruits histone deacetylases in mammalian cells", J BIOL CHEM, 276(48), 2001, pp. 45168-45174

Abstract

Regulated gene expression will provide important platforms from which genefunctions can be investigated and safer means of gene therapy may be developed. Histone deacetylases have recently been shown to play an important role in regulating gene expression. Here we investigated whether a more tightly controlled expression could be achieved by using a novel chimeric repressor that recruits histone deacetylases to a tetracycline-responsive promoter. This chimeric repressor was engineered by fusing the tetracycline repressor (TetR) with an mSin3-interacting domain of human Mad1 and was shown to bind the tetO(2) element with high affinity, and its binding was efficiently abrogated by doxycycline. The chimeric repressor was shown to directly interact with mSin3 of the historic deacetylase complex. This inducible system was further simplified by using a single vector that contained both a chimeric repressor expression cassette and a tetracycline-responsive promoter. When transiently introduced into mammalian cells, the chimeric repressor system exhibited a significantly lower basal level of luciferase activity (up to 25-fold) than that of the TetR control. When stably transfected into HEK 293 cells, the chimeric repressor system was shown to exert a tight control of green fluorescent protein expression in a doxycycline dose- and time-dependent fashion. Therefore, this novel chimeric repressor provides an effective means for more tightly regulated gene expression, and the simplified inducible system may be used for a broad range of basic and clinical studies.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/01/20 alle ore 21:24:21