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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents

ID: 127827.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
A large-scale, gene-driven mutagenesis approach for the functional analysis of the mouse genome
Authors:Hansen, Jens; Floss, Thomas; Van Sloun, Petra; Fuchtbauer, Ernst-Martin; Vauti, Franz; Arnold, Hans-Hennig; Schnutgen, Frank; Wurst, Wolfgang; von Melchner, Harald; Ruiz, Patricia
Date of Publication (YYYY-MM-DD):2003-08-19
Title of Journal:Proceedings of the National Academy of Sciences of the United States of America
Journal Abbrev.:Proc. Natl. Acad. Sci. U. S. A.
Issue / Number:17
Start Page:9918
End Page:9922
Copyright:Copyright © 2003 by the National Academy of Sciences
Review Status:not specified
Audience:Experts Only
Abstract / Description: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; 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.
Comment of the Author/Creator:Published online before print August 6, 2003,
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:GSF, Natl Res Ctr Environm & Hlth, Inst Dev Genet, D-85764 Neuherberg, Germany.; Univ Frankfurt, Sch Med, Lab Mol Hematol, D-60590 Frankfurt, Germany.; Max Planck Inst Immunbiol, Dept Dev Biol, D-79108 Freiburg, Germany.; TU Braunschweig, Dept Cell & Mol Biol, Inst Biochem & Biotechnol, D-38106 Braunschweig, Germany.; Max Planck Inst Psychiat, Dept Mol Neurogenet, D-80804 Munich, Germany.; Max Planck Inst Mol Genet, Dept Vertebrate Genom, D-14195 Berlin, Germany.
Identifiers:ISI:000184926000054 [ID No:1]
ISSN:0027-8424 [ID No:2]
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