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GENETICS
A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome
** 
*Institute of Developmental Genetics,
GSF-National Research Center for Environment and Health, D-85764 Neuherberg,
Germany; Laboratory for Molecular Hematology,
University of Frankfurt Medical School, D-60590 Frankfurt am Main, Germany;
Department of Developmental Biology, Max
Planck Institute of Immunobiology, D-79108 Freiburg, Germany;
¶Department of Cell and Molecular Biology,
Institute of Biochemistry and Biotechnology, TU Braunschweig, D-38106
Braunschweig, Germany; ||Department for Molecular
Neurogenetics, Max Planck Institute of Psychiatry, D-80804 Munich, Germany;
and Department of Vertebrate Genomics,
Max Planck Institute for Molecular Genetics, D-14195 Berlin, Germany
Communicated by Sherman M. Weissman, Yale University School of Medicine, New Haven, CT, May 30, 2003 (received for review November 5, 2002)
A major challenge of the postgenomic era is the functional characterization of every single gene within the mammalian genome. In an effort to address this challenge, we assembled a collection of mutations in mouse embryonic stem (ES) cells, which is the largest publicly accessible collection of such mutations to date. Using four different gene-trap vectors, we generated 5,142 sequences adjacent to the gene-trap integration sites (gene-trap sequence tags; http://genetrap.de) from >11,000 ES cell clones. Although most of the gene-trap vector insertions occurred randomly throughout the genome, we found both vector-independent and vector-specific integration "hot spots." Because >50% of the hot spots were vector-specific, we conclude that the most effective way to saturate the mouse genome with gene-trap insertions is by using a combination of gene-trap vectors. When a random sample of gene-trap integrations was passaged to the germ line, 59% (17 of 29) produced an observable phenotype in transgenic mice, a frequency similar to that achieved by conventional gene targeting. Thus, gene trapping allows a large-scale and cost-effective production of ES cell clones with mutations distributed throughout the genome, a resource likely to accelerate genome annotation and the in vivo modeling of human disease.
Present address: Institute of Molecular and Structural Biology, Aarhus
University, C. F. Mollers Alle, 8000 Aarhus C, Denmark.
** W.W., H.v.M., and P.R. contributed equally to this work.
To whom correspondence should be addressed. E-mail:
ruiz{at}molgen.mpg.de.
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