Published online 19 January 2005
Methods Online |
Gene trap and gene inversion methods for conditional gene inactivation in the mouse
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University Ithaca, NY 14853, USA 1 Computational Biology Service Unit, Cornell Theory Center, Cornell University Ithaca, NY 14853, USA 2 Department of Cancer Biology, Vanderbilt University School of Medicine Nashville, TN 37235, USA
*To whom correspondence should be addressed at Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, T4018 VRT, Box 11, Ithaca, NY 14853-6401, USA. Tel: +1 607 253 3336; Fax: +1 607 253 3317; Email: mik7{at}cornell.edu
Received November 18, 2004. Revised December 23, 2004. Accepted December 23, 2004.
Conditional inactivation of individual genes in mice using site-specific recombinases is an extremely powerful method for determining the complex roles of mammalian genes in developmental and tissue-specific contexts, a major goal of post-genomic research. However, the process of generating mice with recombinase recognition sequences placed at specific locations within a gene, while maintaining a functional allele, is time consuming, expensive and technically challenging. We describe a system that combines gene trap and site-specific DNA inversion to generate mouse embryonic stem (ES) cell clones for the rapid production of conditional knockout mice, and the use of this system in an initial gene trap screen. Gene trapping should allow the selection of thousands of ES cell clones with defined insertions that can be used to generate conditional knockout mice, thereby providing extensive parallelism that eliminates the time-consuming steps of targeting vector construction and homologous recombination for each gene.
Correspondence may also be addressed to Hong-Bo Xin. Tel: +1 607 253 4321; Fax: +1 607 253 3317; Email: hx25{at}cornell.edu