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

A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome
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.
- Max Planck Institute of Neurobiology Germany
- Max Planck Society Germany
- National Center for Health Research United States
- Institute of Genetics and Developmental Biology China (People's Republic of)
Base Sequence, Stem Cells, Genetic Vectors, Mice, Transgenic, DNA, Genomics, Cell Line, Mice, Inbred C57BL, Mice, Mutagenesis, Insertional, Phenotype, Animals, Humans, Sequence Tagged Sites
Base Sequence, Stem Cells, Genetic Vectors, Mice, Transgenic, DNA, Genomics, Cell Line, Mice, Inbred C57BL, Mice, Mutagenesis, Insertional, Phenotype, Animals, Humans, Sequence Tagged Sites
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