Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells
Autor: | Mansour Karimi, Ilse De Vos, Pierre Hilson, Lieven Haenebalcke, Agnieszka Gembarska, Steven Goossens, Bram De Craene, Sarah De Clercq, András D Nagy, Jean-Christophe Marine, Benjamin Drogat, Sonia Bartunkova, Jody J. Haigh, Omar Nyabi, Geert Berx, Michael Naessens, Marion M. Maetens, Katharina Haigh |
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Jazyk: | angličtina |
Rok vydání: | 2009 |
Předmět: |
Genetically modified mouse
Génétique du développement Génétique moléculaire RNA Untranslated Transgene Biochimie Acides nucléiques synthèse des protéines Gene Targeting -- methods Genetic Vectors Biotechnologie Biologie générale Sciences de la matière vivante Cloning Molecular -- methods Mice Transgenic Computational biology Biology Hybrid Cells Cell Line Proteins -- genetics Mice Complementary DNA Genetics Animals Transgenes Cloning Molecular Gene Embryonic Stem Cells Alleles Cloning Recombination Genetic Histologie Gene targeting Proteins Biologie moléculaire Embryonic stem cell Diploidy Transgenesis Cancérologie Génétique cytogénétique Gene Targeting Methods Online Biologie cellulaire Génétique cellulaire Biologie Embryonic Stem Cells -- metabolism Embryologie [animale] |
Zdroj: | Nucleic acids research, 37 (7 Nucleic Acids Research |
Popis: | 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 3-4 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. Journal Article Research Support, Non-U.S. Gov't info:eu-repo/semantics/published |
Databáze: | OpenAIRE |
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