Published online 13 February 2006
Methods Online |
High-throughput trapping of secretory pathway genes in mouse embryonic stem cells
Department of Molecular Hematology, University of Frankfurt Medical School Frankfurt am Main, Germany 1Institute of Developmental Genetics, GSF-National Research Center for Environment and Health Neuherberg, Germany 2Center for Cardiovascular Research, Charité Universitätsmedizin Berlin, Germany 3Department of Vertebrate Genomics, Max-Planck Institute for Molecular Genetics Berlin, Germany 4Department for Molecular Neurogenetics, Max-Planck Institute of Psychiatry Munich, Germany
*To whom correspondence should be addressed. Tel: +49 69 63016696; Fax: +49 69 63016390; Email: melchner{at}em.uni-frankfurt.de
Received December 23, 2005. Revised January 27, 2006. Accepted January 27, 2006.
High-throughput gene trapping is a random approach for inducing insertional mutations across the mouse genome. This approach uses gene trap vectors that simultaneously inactivate and report the expression of the trapped gene at the insertion site, and provide a DNA tag for the rapid identification of the disrupted gene. Gene trapping has been used by both public and private institutions to produce libraries of embryonic stem (ES) cells harboring mutations in single genes. Presently, 
66% of the protein coding genes in the mouse genome have been disrupted by gene trap insertions. Among these, however, genes encoding signal peptides or transmembrane domains (secretory genes) are underrepresented because they are not susceptible to conventional trapping methods. Here, we describe a high-throughput gene trapping strategy that effectively targets secretory genes. We used this strategy to assemble a library of ES cells harboring mutations in 716 unique secretory genes, of which 61% were not trapped by conventional trapping, indicating that the two strategies are complementary. The trapped ES cell lines, which can be ordered from the International Gene Trap Consortium (http://www.genetrap.org), are freely available to the scientific community.
The German Gene Trap Consortium: Silke De-Zolt, Frank Schnütgen, Claudia Seisenberger, Jens Hansen, Melanie Hollatz, Thomas Floss, Patricia Ruiz, Wolfgang Wurst and Harald von Melchner
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