Functional Analysis of Mammalian Genes by a Large Scale Gene Trap Approach in Mouse Embryonic Stem Cells

Thomas Floss
GSF Institute for Environment and Health
Institute of Mammalian Genetics
Ingolstaedter Landstr. 1
85764 Oberschleissheim
telephone: 089-3187-2214
fax: 089-3187-3099
email: tfloss@gsf.de
prestype: Poster
presenter: Thomas Floss

Thomas Floss1, Franz Vauti2, Michael Wiles3, Ernst-Martin Füchtbauer4, Hans-Henning Arnold2, Harald von Melchner5, Jürgen Otte5, Thomas Metz1 and Wolfgang Wurst1

1GSF - National Research Center for Environment and Health, Institute of Mammalian Genetics, Ingolstaedter Landstrasse 1, D-85764 Neuherberg;
2
Department of Cell- and Molecular Biology, Institute of Biochemistry and Biotechnology, TU Braunschweig, Spielmannstr. 7, D-38106 Braunschweig;
3Max-Planck-Institute for Molecular Genetics, Ihnestr. 73, D-14195 Berlin;
4Max-Planck-Institute of Immunobiology, Department of Developmental Biology, Stübeweg 51, D-79108 Freiburg;
5Laboratory for Molecular Hematology, University of Frankfurt Medical School, Theodor-Stern Kai 7, D-60590 Frankfurt am Main.

We have established a research center within the German HUGO project in order to perform a large scale functional analysis of mammalian genes taking advantage of the gene trap technology in combination with mouse embryonic stem (ES) cells. The gene trap technology provides an important tool of the human genome project for the identification and functional characterization of mammalian genes.

We are currently performing a large scale insertional mutagenesis screen in mouse ES cells. The mutagenic events are based on gene trap vector integrations into genes expressed in ES cells, generating a large number of mutant ES clones. The mutated genes are identified using RACE-PCR strategies allowing us to establish an archive of mutated genes generated in ES cells. We will establish mutant mouse lines by germ line transmission in case the genes mutated relate to human diseases.

At present, we have established 14.000 individual, mutated ES cell clones from which 6800 trapped genes have been sequenced (49%). Of the 3848 good quality sequences obtained, 1818 (47%) show homology to relevant genes. Another 903 sequences (23%) have been identified as EST´s and the remaining 1127 sequences (29%) show no homologies to known sequences present in the NCBI-GenBank database. The individual data for each clone, i.e. sequence, expression pattern and eventually mutant phenotype is stored in a public database which is freely accessible to the scientific community (http://tikus.gsf.de). These data will extend the international EST program and will contribute to unravel gene function genome wide.



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