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Generation of mice with a 200-kb amyloid precursor protein gene deletion by Cre recombinase-mediated site-specific recombination in embryonic stem cells
- Source :
- Proceedings of the National Academy of Sciences of the United States of America. 93(12)
- Publication Year :
- 1996
-
Abstract
- Gene disruptions and deletions of up to 20kb have been generated by homologous recombination with appropriate targeting vectors in murine embryonic stem (ES) cells. Because we could not obtain a deletion of about 200 kb in the mouse amyloid precursor protein gene by the classical technique, we employed strategies involving the insertion of loxP sites upstream and downstream of the region to be deleted by homologous recombination and elicited excision of the loxP-flanked region by introduction of a Cre expression vector into the ES cells. In the first approach, the loxP sequences were inserted in two successive steps and after each step, ES cell clones were isolated and characterized. Deletion of the loxP-flanked sequence was accomplished by introducing the cre gene in a third step. In the second approach, ES cells containing the upstream loxP cassette were electroporated simultaneously with the downstream loxP targeting vector and the Cre expression plasmid. ES cells were obtained that gave rise to chimeric mice capable of germ-line transmission of the deleted amyloid precursor protein allele.
- Subjects :
- Molecular Sequence Data
Cre recombinase
Biology
Amyloid beta-Protein Precursor
Mice
Viral Proteins
Amyloid precursor protein
Animals
Site-specific recombinase technology
Site-specific recombination
Amino Acid Sequence
Floxing
DNA Primers
Recombination, Genetic
Multidisciplinary
Expression vector
Base Sequence
Integrases
Stem Cells
Embryo, Mammalian
Molecular biology
Mice, Mutant Strains
DNA Nucleotidyltransferases
biology.protein
Stem cell
Homologous recombination
Gene Deletion
Research Article
Subjects
Details
- ISSN :
- 00278424
- Volume :
- 93
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Accession number :
- edsair.doi.dedup.....64ca4aa31bca1606e948b4a49d5cbc9f