caa a22 4500 445823143 CHVBK 20180317145250.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10616-011-9335-z doi (NATIONALLICENCE)springer-10.1007/s10616-011-9335-z Development of a tightly-regulated tetracycline-dependent transcriptional activator and repressor co-expression system for the strong induction of transgene expression [Elektronische Daten] [Hiroshi Hosoda, Takahisa Miyao, Shusaku Uchida, Shinsuke Sakai, Satoshi Kida] The teteracycline (Tc)-dependent and -inducible transcriptional activator (rtTA) system has been used to express regulated transgene expression in vitro and in vivo. However, previous reports have demonstrated that, even in the absence of Tc, the rtTA binds weakly to the tetracycline response element (TRE), leading to a low level of background activity. In order to reduce the leaky gene expression induced by rtTA, we previously established a tightly regulated system (A-IRES-R system) that makes use of both the rtTA (A) and a Tc-dependent repressor (TetR-Kruppel-associated box; KRAB) (R). In addition, others have described a transactivator rtTA2-M2 (M2) that displays higher sensitivity to Dox than rtTA. In this study, to further develop the A-IRES-R system, we generated a derivative Tc system (M2-IRES-R system) that co-expresses both rtTA-M2 and TetR-KRAB from a single vector. We show that compared to the A-IRES-R system, the M2-IRES-R system leads to a greater level of induced TRE-mediated transcription in the presence of doxycycline (Dox) and yet displays a similar level of basal TRE-mediated transcription in the absence of Dox. Furthermore, the M2-IRES-R system also displays less leaky gene expression in the absence of Dox compared to rtTA-M2 and rtTA systems. Taken together, our results suggest that the M2-IRES-R system enables to tightly regulate and highly induce the expression of transgene compared to other systems. Springer Science+Business Media B.V., 2011 rtTA-M2 nationallicence Tight regulation nationallicence Transgenic technique nationallicence Basal transcription nationallicence Hosoda Hiroshi Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut Miyao Takahisa Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut Uchida Shusaku Department of Agricultural Chemistry, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut Sakai Shinsuke Department of Agricultural Chemistry, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut Kida Satoshi Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut Cytotechnology Springer Netherlands 63/3(2011-05-01), 211-216 0920-9069 63:3<211 2011 63 10616 https://doi.org/10.1007/s10616-011-9335-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9335-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hosoda Hiroshi Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut NATIONALLICENCE 700 1- Miyao Takahisa Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut NATIONALLICENCE 700 1- Uchida Shusaku Department of Agricultural Chemistry, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut NATIONALLICENCE 700 1- Sakai Shinsuke Department of Agricultural Chemistry, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut NATIONALLICENCE 700 1- Kida Satoshi Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, 156-8502, Tokyo, Japan aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/3(2011-05-01), 211-216 0920-9069 63:3<211 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer