Genomewide Production of Multipurpose Alleles for the Functional Analysis of the Mouse Genome

A type of retroviral gene trap vectors has been developed that can induce conditional mutations in most genes expressed in mouse embryonic stem (ES) cells. The vectors rely on directional site-specific recombination systems that can repair and reinduce gene trap mutations when activated in successio...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 102; no. 20; pp. 7221 - 7226
Main Authors Schnütgen, Frank, De-Zolt, Silke, Van Sloun, Petra, Hollatz, Melanie, Floss, Thomas, Hansen, Jens, Altschmied, Joachim, Seisenberger, Claudia, Ghyselinck, Norbert B., Ruiz, Patricia, Chambon, Pierre, Wurst, Wolfgang, von Melchner, Harald
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 17.05.2005
National Acad Sciences
Subjects
Alleles
Analysis
Animals
Biochemistry, Molecular Biology
Biological Sciences
Blotting, Northern
Blotting, Western
Cell Line
Cell lines
Computational Biology
Embryo
Embryo, Mammalian - cytology
Genes
Genes - genetics
Genetic mutation
Genetic Vectors
Genetic Vectors - genetics
Genome
Genomes
Genomics
Genomics - methods
Introns
Life Sciences
Mice
Mice - genetics
Molecular biology
Mutagenesis
Mutagenesis, Site-Directed
Mutation
Mutation - genetics
Plasmids
Polymerase Chain Reaction
Retroviridae
Rodents
Somatic cells
Stem Cells
Stem Cells - cytology
Stem Cells - metabolism
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Summary:A type of retroviral gene trap vectors has been developed that can induce conditional mutations in most genes expressed in mouse embryonic stem (ES) cells. The vectors rely on directional site-specific recombination systems that can repair and reinduce gene trap mutations when activated in succession. After the gene traps are inserted into the mouse genome, genetic mutations can be produced at a particular time and place in somatic cells. In addition to their conditional features, the vectors create multipurpose alleles amenable to a wide range of postinsertional modifications. Here we have used these directional recombination vectors to assemble the largest library of ES cell lines with conditional mutations in single genes yet assembled, presently totaling 1,000 unique genes. The trapped ES cell lines, which can be ordered from the German Gene Trap Consortium, are freely available to the scientific community.
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PMCID: PMC1129123
Contributed by Pierre Chambon, March 22, 2005
Abbreviations: GTST, gene trap sequence tag; SA, splice acceptor; RTs, recombinase target sequences; pA, polyadenylation sequence; FlEx, flip-excision; RBBP7, retinoblastoma binding protein 7; Glt28d1, glycosyltransferase 28 domain containing 1 gene.
To whom correspondence should be addressed at: Department of Molecular Hematology, University of Frankfurt Medical School, Theodor-Stern-Kai 7, 60590 Frankfurt am Main, Germany. E-mail: melchner@em.uni-frankfurt.de.
Author contributions: F.S., S.D.-Z., P.V.S., N.B.G., P.C., W.W., and H.v.M. designed research; F.S., S.D.-Z., P.V.S., M.H., T.F., J.A., C.S., and P.R. performed research; F.S., S.D.-Z., P.V.S., M.H., T.F., P.R., and H.v.M. contributed new reagents/analytic tools; F.S., S.D.-Z., P.V.S., M.H., T.F., J.H., P.R., P.C., W.W., and H.v.M. analyzed data; and H.v.M. wrote the paper.
F.S., S.D.-Z., P.V.S., M.H., T.F., J.H., C.S., P.R., W.W., and H.v.M. are members of The German Gene Trap Consortium.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0502273102