title: "Tet-On" system toward hepatic differentiation of human mesenchymal stem cells by hepatocyte nuclear factor.
abstract: "Tet-On" system requires two DNA constructs: the first one is a transcription regulatory unit, rtTA and the second construct is the responsive element Escherichia coli sequences (tetO) linked to Pcmv driven target gene. In the absence of inducing agent doxycycline (Dox), a tetracycline derivative, rtTA does not bind to or binds weakly to operator sequences of tetO; therefore, no target gene is transcribed. However, in the presence of Dox, tTA binds to tetO and pcmv, which in turn activates the target gene. In general, the induction of transgene by Dox is rapid and can occur within hours in some systems, offering an advantage over the original tTA system for studying acute effects of transgenes. Recently, we have established a Tet-regulated expression system for hepatocyte nuclear factor 3β (HNF3β) to investigate the potency of hepatic differentiation of human mesenchymal stem cells (MSC) by HNF3β.
authors.affiliation: Division of Molecular and Genetic Medicine, Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medicine, Tottori University, Yonago, Japan. gshiota@med.tottori-u.ac.jp
authors.forename: Goshi
authors.initials: G
authors.lastname: Shiota
authors.forename: Yoko
authors.initials: Y
authors.lastname: Yoshida
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004318:Doxycycline; D051542:Hepatocyte Nuclear Factor 3-beta / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015533:Transcriptional Activation / Q000235:genetics / Q000502:physiology; D019076:Transgenes / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; C083139:Tet O resistance protein, Bacteria; D051542:Hepatocyte Nuclear Factor 3-beta; D004318:Doxycycline