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Site-directed integration of the cre gene mediated by Cre recombinase using a combination of mutant lox sites.
- Source :
-
Nucleic acids research [Nucleic Acids Res] 2002 Oct 01; Vol. 30 (19), pp. e103. - Publication Year :
- 2002
-
Abstract
- The Cre-lox system is an important tool for genetic manipulation. To promote integrative reactions, two strategies using mutant lox sites have been developed. One is the left element/right element (LE/RE)-mutant strategy and the other is the cassette exchange strategy using heterospecific lox sites such as lox511 or lox2272. We compared the recombination efficiencies using these mutant lox sites in embryonic stem (ES) cells, and found that the combination of the LE/RE mutant and lox2272 showed high recombination efficiency and stability of the recombined structure. Taking advantage of this stability, we successfully integrated the cre gene into the mutant lox sites by Cre-mediated recombination. Germ line chimeric mice were produced from the cre-integrated ES cell clones, and Cre-expressing mouse lines were established. The inserted cre gene was stably maintained through the generations. This cre knock-in system using the LE/RE-lox2272 combination should be useful for the production of various cre mice for gene targeting or gene trapping.
- Subjects :
- Animals
Base Sequence
Binding Sites genetics
Cell Line
Female
Integrases metabolism
Male
Mice
Mice, Inbred C57BL
Mice, Inbred Strains
Mice, Transgenic
Mutagenesis, Site-Directed
Mutation
Plasmids genetics
Viral Proteins metabolism
Integrases genetics
Recombination, Genetic
Viral Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1362-4962
- Volume :
- 30
- Issue :
- 19
- Database :
- MEDLINE
- Journal :
- Nucleic acids research
- Publication Type :
- Academic Journal
- Accession number :
- 12364620
- Full Text :
- https://doi.org/10.1093/nar/gnf102