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Gene-Trap Vectors and Mutagenesis

  • Silke De-Zolt
  • Joachim Altschmied
  • Patricia Ruiz
  • Harald von Melchner
  • Frank Schnütgen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 530)

Abstract

Gene trapping can be used to introduce insertional mutations into the genome of mouse embryonic stem cells (ESCs). The method has been adapted for high-throughput use, in an effort to inactivate all genes in the mouse genome. Gene trapping is performed with vectors that simultaneously inactivate and report the expression of the trapped gene and provide a molecular tag for its rapid identification. Gene-trap approaches have been used successfully in the past by both academic and commercial organizations to create libraries of ESC lines harboring mutations in single genes that can be used for making mice. Presently, approximately 70% of the protein-coding genes in the mouse genome have been disrupted by gene-trap insertions. Here we describe the basic methodology used to induce and characterize gene-trap mutations in ESCs.

Key words

High-throughput mutagenesis gene trapping ES cells 

Notes

Acknowledgements

We thank all members of the von Melchner laboratory for helpful discussions and suggestions. This work was supported by grants from the Bundesministerium für Bildung und Forschung (BMBF) to the German Gene Trap Consortium and from the Deutsche Forschungsgemeinschaft (DFG) to HvM.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Silke De-Zolt
    • 1
  • Joachim Altschmied
    • 1
  • Patricia Ruiz
    • 2
  • Harald von Melchner
    • 1
  • Frank Schnütgen
    • 1
  1. 1.Department of Molecular HematologyUniversity of FrankfurtFrankfurt am MainGermany
  2. 2.Center for Cardiovascular ResearchCharité – Universitätsmedizin BerlinGermany

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