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Titolo:
A model system for studying postnatal myogenesis with tetracycline-responsive, genetically engineered clonal myoblasts in vitro and in vivo
Autore:
Link, D; Irintchev, A; Knauf, U; Wernig, A; Starzinski-Powitz, A;
Indirizzi:
Xantos Biomed AG, D-82152 Martinsried, Germany Xantos Biomed AG Martinsried Germany D-82152 -82152 Martinsried, Germany Univ Bonn, Dept Physiol, D-53111 Bonn, Germany Univ Bonn Bonn Germany D-53111 Bonn, Dept Physiol, D-53111 Bonn, Germany
Titolo Testata:
EXPERIMENTAL CELL RESEARCH
fascicolo: 2, volume: 270, anno: 2001,
pagine: 138 - 150
SICI:
0014-4827(20011101)270:2<138:AMSFSP>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
INDUCIBLE GENE-EXPRESSION; HIGHER EUKARYOTIC CELLS; SKELETAL-MUSCLE CELLS; TRANSGENIC MICE; MAMMALIAN-CELLS; MOUSE MUSCLES; MDX MICE; DIFFERENTIATION; IMPLANTATION; TRANSPLANTATION;
Keywords:
skeletal muscle; myoblasts; satellite cells; transplantation; Tet-off system; luciferase; tetracycline; tTA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Starzinski-Powitz, A Univ Frankfurt, Dept Anthropol & Human Genet, Siesmayerstr 70, D-60054 Frankfurt, Germany Univ Frankfurt Siesmayerstr 70 Frankfurt Germany D-60054
Citazione:
D. Link et al., "A model system for studying postnatal myogenesis with tetracycline-responsive, genetically engineered clonal myoblasts in vitro and in vivo", EXP CELL RE, 270(2), 2001, pp. 138-150

Abstract

The aim of this work was to introduce a tetracycline-responsive (Tet-off) gene expression system into myoblasts in order to regulate a reporter gene not only in vitro but also particularly in muscles implanted with these engineered myoblasts. Mouse myoblasts from a long-term culture (i28 cells) were transfected initially to generate and characterize two stable master clones expressing tetracycline-responsive transactivator protein tTA. Like parental i28 myoblasts, these clones differentiated well in vitro. The second step introduced the firefly (Photinus pyralis) luciferase gene into one of the stable tTA clones producing double transfectants expressing luciferase in the absence of tetracycline. Addition of tetracycline (1 mug ml(-1)) resulted in at least 100-fold decreases in luciferase activity within 8 h in both growing and differentiating myoblast cultures. Enzyme activity was rapidly restored after tetracycline was removed (8 h). After successful implantation of these myoblasts into damaged mouse muscles, luciferase expression in the matured progeny cells could be regulated by oral application of doxycycline for at least 1 month. The tetracycline-responsive master clones are potentially powerful tools for studying the function of various genes in postnatal myogenesis. (C) 2001 Academic Press.

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Documento generato il 13/07/20 alle ore 14:53:16