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Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells

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Abstract
The ability to rapidly and efficiently generate reliable Cre/loxP conditional transgenic mice would greatly complement global high-throughput gene targeting initiatives aimed at identifying gene function in the mouse. We report here the generation of Cre/loxP conditional ROSA26-targeted ES cells within 34 weeks by using Gateway cloning to build the target vectors. The cDNA of the gene of interest can be expressed either directly by the ROSA26 promoter providing a moderate level of expression or by a CAGG promoter placed in the ROSA26 locus providing higher transgene expression. Utilization of F1 hybrid ES cells with exceptional developmental potential allows the production of germ line transmitting, fully or highly ES cell-derived mice by aggregation of cells with diploid embryos. The presented streamlined procedures accelerate the examination of phenotypical consequences of transgene expression. It also provides a unique tool for comparing the biological activity of polymorphic or splice variants of a gene, or products of different genes functioning in the same or parallel pathways in an overlapping manner.

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Chicago
Nyabi, Omar, Michaël Naessens, Katharina Haigh, Agnieszka Gembarska, Steven Goossens, Marion Maetens, Sarah De Clercq, et al. 2009. “Efficient Mouse Transgenesis Using Gateway-compatible ROSA26 Locus Targeting Vectors and F1 Hybrid ES Cells.” Nucleic Acids Research 37 (7): e55.
APA
Nyabi, O., Naessens, M., Haigh, K., Gembarska, A., Goossens, S., Maetens, M., De Clercq, S., et al. (2009). Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells. NUCLEIC ACIDS RESEARCH, 37(7), e55.
Vancouver
1.
Nyabi O, Naessens M, Haigh K, Gembarska A, Goossens S, Maetens M, et al. Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells. NUCLEIC ACIDS RESEARCH. 2009;37(7):e55.
MLA
Nyabi, Omar, Michaël Naessens, Katharina Haigh, et al. “Efficient Mouse Transgenesis Using Gateway-compatible ROSA26 Locus Targeting Vectors and F1 Hybrid ES Cells.” NUCLEIC ACIDS RESEARCH 37.7 (2009): e55. Print.
@article{665456,
  abstract     = {The ability to rapidly and efficiently generate reliable Cre/loxP conditional transgenic mice would greatly complement global high-throughput gene targeting initiatives aimed at identifying gene function in the mouse. We report here the generation of Cre/loxP conditional ROSA26-targeted ES cells within 34 weeks by using Gateway cloning to build the target vectors. The cDNA of the gene of interest can be expressed either directly by the ROSA26 promoter providing a moderate level of expression or by a CAGG promoter placed in the ROSA26 locus providing higher transgene expression. Utilization of F1 hybrid ES cells with exceptional developmental potential allows the production of germ line transmitting, fully or highly ES cell-derived mice by aggregation of cells with diploid embryos. The presented streamlined procedures accelerate the examination of phenotypical consequences of transgene expression. It also provides a unique tool for comparing the biological activity of polymorphic or splice variants of a gene, or products of different genes functioning in the same or parallel pathways in an overlapping manner.},
  author       = {Nyabi, Omar and Naessens, Micha{\"e}l and Haigh, Katharina and Gembarska, Agnieszka and Goossens, Steven and Maetens, Marion and De Clercq, Sarah and Drogat, Benjamin and Haenebalcke, Lieven and Bartunkova, Sona and De Vos, Ilse and De Craene, Bram and Karimi, Mansour and Berx, Geert and Nagy, Andras and Hilson, Pierre and Marine, Jean-Christophe and Haigh, Jody},
  issn         = {0305-1048},
  journal      = {NUCLEIC ACIDS RESEARCH},
  language     = {eng},
  number       = {7},
  title        = {Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells},
  url          = {http://dx.doi.org/10.1093/nar/gkp112},
  volume       = {37},
  year         = {2009},
}

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