Generating conditional knockout mice
- PMID: 21080282
- DOI: 10.1007/978-1-60761-974-1_12
Generating conditional knockout mice
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.
Similar articles
-
Conditional gene targeting in the mouse nervous system: Insights into brain function and diseases.Pharmacol Ther. 2007 Mar;113(3):619-34. doi: 10.1016/j.pharmthera.2006.12.003. Epub 2007 Jan 10. Pharmacol Ther. 2007. PMID: 17289150 Review.
-
Construction of engineered murine embryonic stem cells with conditional knockout of FGFR2 depending on Cre-loxP.Biocell. 2006 Aug;30(2):269-78. Biocell. 2006. PMID: 16972551
-
Site- and time-specific gene targeting in the mouse.Methods. 2001 May;24(1):71-80. doi: 10.1006/meth.2001.1159. Methods. 2001. PMID: 11327805
-
A directional strategy for monitoring Cre-mediated recombination at the cellular level in the mouse.Nat Biotechnol. 2003 May;21(5):562-5. doi: 10.1038/nbt811. Epub 2003 Mar 31. Nat Biotechnol. 2003. PMID: 12665802
-
Inducible gene targeting in mice using the Cre/lox system.Methods. 1998 Apr;14(4):381-92. doi: 10.1006/meth.1998.0593. Methods. 1998. PMID: 9608509 Review.
Cited by
-
Lethal adulthood myelin breakdown by oligodendrocyte-specific Ddx54 knockout.iScience. 2023 Jul 21;26(10):107448. doi: 10.1016/j.isci.2023.107448. eCollection 2023 Oct 20. iScience. 2023. PMID: 37720086 Free PMC article.
-
Targeted Disruption of the MORG1 Gene in Mice Causes Embryonic Resorption in Early Phase of Development.Biomolecules. 2023 Jun 24;13(7):1037. doi: 10.3390/biom13071037. Biomolecules. 2023. PMID: 37509073 Free PMC article.
-
Methodological aspects of in vivo axial loading in rodents: a systematic review.J Musculoskelet Neuronal Interact. 2023 Jun 1;23(2):236-262. J Musculoskelet Neuronal Interact. 2023. PMID: 37259664 Free PMC article.
-
Defining Gene Function in Spermatogonial Stem Cells Through Conditional Knockout Approaches.Methods Mol Biol. 2023;2656:261-307. doi: 10.1007/978-1-0716-3139-3_15. Methods Mol Biol. 2023. PMID: 37249877
-
Knockout of the Complex III subunit Uqcrh causes bioenergetic impairment and cardiac contractile dysfunction.Mamm Genome. 2023 Jun;34(2):229-243. doi: 10.1007/s00335-022-09973-w. Epub 2022 Dec 24. Mamm Genome. 2023. PMID: 36565314 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources