International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany


POSTER 55 - FUNCTIONAL ANALYSIS OF THE MAMMALIAN GENOME BY LARGE SCALE GENE TRAP MUTAGENESIS

Ruiz P
Department of Vertebrate Genomics, Max-Planck Institute for Molecular Genetics, Berlin, Germany

Co-Authors: 2) Hansen J, 2) Floss T, 3) van Sloun P, 3) Schnütgen F (3), 4) Füchtbauer EM, 5) Vauti F, 1) Lehrach H, 3) von Melchner H, 2) and Wurst W
Institutions: 1) Department of Vertebrate Genomics, Max-Planck Institute for Molecular Genetics, Berlin, Germany; 2) Institute of Developmental Genetics, GSF-National Research Center for Environment and Health, Neuherberg, Germany; 3) Laboratory for Molecular Hematology, University of Frankfurt Medical School, Frankfurt am Main, Germany; 4) Department of Developmental Biology, Max-Planck Institute of Immunobiology, Freiburg, Germany; 5) Department of Cell and Molecular Biology, Institute of Biochemistry and Biotechnology, TU Braunschweig, Braunschweig, Germany

We have established a Research Consortium (German Gene trap Consortium, GGTC) to carry out large-scale gene trap mutagenesis in ES cells. Its goal is to contribute to the saturation mutagenesis of the mouse genome and to generate a mouse model for each gene in cooperation with the International Mouse Mutant Consortium (IMMC). Towards this goal, the GGTC has generated 11,266 mutant ES cell lines and has identified the gene trap integration sites in 8423 clones. Of the generated gene trap sequence tags (GTSTs), 5142 informative sequences were obtained of which 3750 corresponded to known genes, 623 to ESTs and 679 to putative novel genes.

We compared the insertion performance of four different gene trap vectors: pT1βgeo, pT1ATGβgeo, U3βgeo and Rosaβgeo. The integration sites of all vectors were found randomly and evenly distributed over the entire mouse genome, although several hot spots were identified for the different vectors. In addition we identified integrations into all different classes of gene products, whereby signal-sequence containing proteins are underrepresented.

Of all integrations into previously characterized genes, 66 occurred in genes involved in human disease. Furthermore, we have generated over 70 germ line chimeras and could show that 62% of resulting homozygous mutants exhibit an obvious phenotype. Finally, data obtained from individual clones, such as GTSTs, expression patterns and phenotypes, are deposited in the GGTC's database which is publicly accessible via http://genetrap.de. Corresponding cell lines, stored frozen at the GSF in Munich, are available freely upon request.


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