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

Omar Nyabi, Michaël Naessens UGent, Katharina Haigh UGent, Agnieszka Gembarska, Steven Goossens UGent, Marion Maetens, Sarah De Clercq UGent, Benjamin Drogat UGent, Lieven Haenebalcke, Sona Bartunkova UGent, et al. (2009) NUCLEIC ACIDS RESEARCH. 37(7).
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
journal title
NUCLEIC ACIDS RESEARCH
Nucleic Acids Res.
volume
37
issue
7
pages
e55 -
Web of Science type
Article
Web of Science id
000265741600038
JCR category
BIOCHEMISTRY & MOLECULAR BIOLOGY
JCR impact factor
7.479 (2009)
JCR rank
27/281 (2009)
JCR quartile
1 (2009)
ISSN
0305-1048
DOI
10.1093/nar/gkp112
language
English
UGent publication?
yes
classification
A1
id
665456
handle
http://hdl.handle.net/1854/LU-665456
date created
2009-05-26 09:02:40
date last changed
2016-12-19 15:46:53
@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},
}

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.