naa a22 4500 510739059 CHVBK 20180411083021.0 cr unu---uuuuu 180411e20131201xx s 000 0 eng 10.1007/s12015-013-9458-z doi (NATIONALLICENCE)springer-10.1007/s12015-013-9458-z Efficient ROSA26-Based Conditional and/or Inducible Transgenesis Using RMCE-Compatible F1 Hybrid Mouse Embryonic Stem Cells [Elektronische Daten] [Lieven Haenebalcke, Steven Goossens, Michael Naessens, Natascha Kruse, Morvarid Ghahremani, Sonia Bartunkova, Katharina Haigh, Tim Pieters, Pieterjan Dierickx, Benjamin Drogat, Omar Nyabi, Dagmar Wirth, Jody Haigh] The conditional Cre/loxP system and/or the doxycycline (Dox) inducible Tet-on/off system are widely used in mouse transgenesis but often require time consuming, inefficient cloning/screening steps and extensive mouse breeding strategies. We have therefore developed a highly efficient Gateway- and recombinase-mediated cassette exchange (RMCE)-compatible system to target conditional and/or inducible constructs to the ROSA26 locus of F1 hybrid Bl6/129 ESCs, called G4 ROSALUC ESCs. By combining the Cre/loxP system with or without the inducible Tet-on system using Gateway cloning, we can rapidly generate spatial and/or temporal controllable gain-of-function constructs that can be targeted to the RMCE-compatible ROSA26 locus of the G4 ROSALUC ESCs with efficiencies close to 100%. These novel ESC-based technologies allow for the creation of multiple gain-of-function conditional and/or inducible transgenic ESC clones and mouse lines in a highly efficient and locus specific manner. Importantly, incorporating insulator sequences into the Dox-inducible vector system resulted in robust, stable transgene expression in undifferentiated ESCs but could not fully overcome transgene mosaicism in the differentiated state. Springer Science+Business Media New York, 2013 Mouse embryonic stem cells nationallicence Gateway technology nationallicence ROSA26 nationallicence Recombinase mediated cassette exchange (RMCE) nationallicence Conditional/Inducible (COIN) nationallicence Insulators nationallicence Haenebalcke Lieven VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Goossens Steven VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Naessens Michael VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Kruse Natascha Helmholtz Centre for Infection Research, Braunschweig, Germany aut Ghahremani Morvarid VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Bartunkova Sonia VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Haigh Katharina VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Pieters Tim VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Dierickx Pieterjan VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Drogat Benjamin VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Nyabi Omar VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Wirth Dagmar Helmholtz Centre for Infection Research, Braunschweig, Germany aut Haigh Jody VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/6(2013-12-01), 774-785 1550-8943 9:6<774 2013 9 12015 https://doi.org/10.1007/s12015-013-9458-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12015-013-9458-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Haenebalcke Lieven VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Goossens Steven VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Naessens Michael VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Kruse Natascha Helmholtz Centre for Infection Research, Braunschweig, Germany aut NATIONALLICENCE 700 1- Ghahremani Morvarid VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Bartunkova Sonia VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Haigh Katharina VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Pieters Tim VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Dierickx Pieterjan VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Drogat Benjamin VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Nyabi Omar VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 700 1- Wirth Dagmar Helmholtz Centre for Infection Research, Braunschweig, Germany aut NATIONALLICENCE 700 1- Haigh Jody VIB Department for Molecular Biomedical Research, Vascular Cell Biology Unit, Ghent, Belgium aut NATIONALLICENCE 773 0- Stem Cell Reviews and Reports Springer US; http://www.springer-ny.com 9/6(2013-12-01), 774-785 1550-8943 9:6<774 2013 9 12015 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer