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Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells

(2013) STEM CELL REVIEWS AND REPORTS. 9(6). p.774-785
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Abstract
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.
Keywords
ROSA26, Recombinase mediated cassette exchange (RMCE), SITE-SPECIFIC INTEGRATION, GENE-EXPRESSION, IN-VIVO, ONCOGENE ADDICTION, RNA INTERFERENCE, CHROMOSOMAL LOCI, ES CELLS, MICE, SYSTEM, Gateway technology, Mouse embryonic stem cells, Insulators, GREEN FLUORESCENT PROTEIN, Conditional/Inducible (COIN)

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Citation

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MLA
Haenebalcke, Lieven, et al. “Efficient ROSA26-Based Conditional and/or Inducible Transgenesis Using RMCE-Compatible F1 Hybrid Mouse Embryonic Stem Cells.” STEM CELL REVIEWS AND REPORTS, vol. 9, no. 6, 2013, pp. 774–85, doi:10.1007/s12015-013-9458-z.
APA
Haenebalcke, L., Goossens, S., Naessens, M., Kruse, N., Farhang Ghahremani, M., Bartunkova, S., … Haigh, J. (2013). Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells. STEM CELL REVIEWS AND REPORTS, 9(6), 774–785. https://doi.org/10.1007/s12015-013-9458-z
Chicago author-date
Haenebalcke, Lieven, Steven Goossens, Michaël Naessens, Natascha Kruse, Morvarid Farhang Ghahremani, Sona Bartunkova, Katharina Haigh, et al. 2013. “Efficient ROSA26-Based Conditional and/or Inducible Transgenesis Using RMCE-Compatible F1 Hybrid Mouse Embryonic Stem Cells.” STEM CELL REVIEWS AND REPORTS 9 (6): 774–85. https://doi.org/10.1007/s12015-013-9458-z.
Chicago author-date (all authors)
Haenebalcke, Lieven, Steven Goossens, Michaël Naessens, Natascha Kruse, Morvarid Farhang Ghahremani, Sona Bartunkova, Katharina Haigh, Tim Pieters, Pieterjan Dierickx, Benjamin Drogat, Omar Nyabi, Dagmar Wirth, and Jody Haigh. 2013. “Efficient ROSA26-Based Conditional and/or Inducible Transgenesis Using RMCE-Compatible F1 Hybrid Mouse Embryonic Stem Cells.” STEM CELL REVIEWS AND REPORTS 9 (6): 774–785. doi:10.1007/s12015-013-9458-z.
Vancouver
1.
Haenebalcke L, Goossens S, Naessens M, Kruse N, Farhang Ghahremani M, Bartunkova S, et al. Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells. STEM CELL REVIEWS AND REPORTS. 2013;9(6):774–85.
IEEE
[1]
L. Haenebalcke et al., “Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells,” STEM CELL REVIEWS AND REPORTS, vol. 9, no. 6, pp. 774–785, 2013.
@article{4213018,
  abstract     = {{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.}},
  author       = {{Haenebalcke, Lieven and Goossens, Steven and Naessens, Michaël and Kruse, Natascha and Farhang Ghahremani, Morvarid and Bartunkova, Sona and Haigh, Katharina and Pieters, Tim and Dierickx, Pieterjan and Drogat, Benjamin and Nyabi, Omar and Wirth, Dagmar and Haigh, Jody}},
  issn         = {{1550-8943}},
  journal      = {{STEM CELL REVIEWS AND REPORTS}},
  keywords     = {{ROSA26,Recombinase mediated cassette exchange (RMCE),SITE-SPECIFIC INTEGRATION,GENE-EXPRESSION,IN-VIVO,ONCOGENE ADDICTION,RNA INTERFERENCE,CHROMOSOMAL LOCI,ES CELLS,MICE,SYSTEM,Gateway technology,Mouse embryonic stem cells,Insulators,GREEN FLUORESCENT PROTEIN,Conditional/Inducible (COIN)}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{774--785}},
  title        = {{Efficient ROSA26-based conditional and/or inducible transgenesis using RMCE-compatible F1 hybrid mouse embryonic stem cells}},
  url          = {{http://dx.doi.org/10.1007/s12015-013-9458-z}},
  volume       = {{9}},
  year         = {{2013}},
}

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