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
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