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A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome

MPG-Autoren

Ruiz,  Patricia
Max Planck Society;

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Zitation

Hansen, J., Floss, T., Van Sloun, P., Fuchtbauer, E.-M., Vauti, F., Arnold, H.-H., et al. (2003). A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome. Proceedings of the National Academy of Sciences of the United States of America, 100(17), 9918-9922.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0010-89CC-E
Zusammenfassung
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.