Generating Conditional Knockout Mice
- 4 October 2010
- book chapter
- Published by Springer Science and Business Media LLC
- Vol. 693, 205-231
- https://doi.org/10.1007/978-1-60761-974-1_12
Abstract
Gene targeting in ES cells is extensively used to generate designed mouse mutants and to study gene function in vivo . Knockout mice that harbor a null allele in their germline provide appropriate genetic models of inherited diseases and often exhibit embryonic or early postnatal lethality. To study gene function in adult mice and in selected cell types, a refined strategy for conditional gene inactivation has been developed that relies on the DNA recombinase Cre and its recognition (loxP) sites. For conditional mutagenesis, a target gene is modified by the insertion of two loxP sites that enable to excise the flanked (floxed) gene segment through Cre-mediated recombination. Conditional mutant mice are obtained by crossing the floxed strain with a Cre transgenic line such that the target gene becomes inactivated in vivo within the expression domain of Cre. A large collection of Cre transgenic lines has been generated over time and can be used in a combinatorial manner to achieve gene inactivation in many different cell types. A growing number of CreER^T2 transgenic mice further allows for inducible inactivation of floxed alleles in adult mice upon administration of tamoxifen. This chapter covers the design and construction of loxP flanked alleles and refers to the vectors, ES cells, and mice generated by the European conditional mouse mutagenesis (EUCOMM) project. We further describe the design and use of Cre and CreER^T2 transgenic mice and a convenient breeding strategy to raise conditional mutants and controls for phenotype analysis.Keywords
This publication has 88 references indexed in Scilit:
- Agouti C57BL/6N embryonic stem cells for mouse genetic resourcesNature Methods, 2009
- Characterization of the Frizzled10‐CreER™ transgenic mouse: An inducible Cre line for the study of Cajal‐Retzius cell developmentgenesis, 2009
- Fate mapping using Cited1-CreERT2 mice demonstrates that the cap mesenchyme contains self-renewing progenitor cells and gives rise exclusively to nephronic epitheliaDevelopmental Biology, 2008
- Vagaries of conditional gene targetingNature Immunology, 2007
- Conditional brain-specific knockdown of MAPK using Cre/loxP regulated RNA interferenceNucleic Acids Research, 2007
- Acute postnatal ablation of Hif-2 α results in anemiaProceedings of the National Academy of Sciences of the United States of America, 2007
- Generation of a transgenic mouse model with chondrocyte‐specific and tamoxifen‐inducible expression of Cre recombinasegenesis, 2007
- Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide–based retroviral vectorNature Biotechnology, 2004
- Inducible Gene Targeting in MiceScience, 1995
- Acre-transgenic mouse strain for the ubiquitous deletion ofloxP-flanked gene segments including deletion in germ cellsNucleic Acids Research, 1995