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Publication : Generation of mice with a 200-kb amyloid precursor protein gene deletion by Cre recombinase-mediated site-specific recombination in embryonic stem cells.

First Author  Li ZW Year  1996
Journal  Proc Natl Acad Sci U S A Volume  93
Issue  12 Pages  6158-62
PubMed ID  8650236 Mgi Jnum  J:36648
Mgi Id  MGI:84075 Doi  10.1073/pnas.93.12.6158
Citation  Li ZW, et al. (1996) Generation of mice with a 200-kb amyloid precursor protein gene deletion by Cre recombinase-mediated site-specific recombination in embryonic stem cells [published erratum appears in Proc Natl Acad Sci U S A 1996 Oct 15;93(21):12052]. Proc Natl Acad Sci U S A 93(12):6158-62
abstractText  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.
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