These NPp53BR2 mice contain the Brca2flox, Rosa26-CAG-LSL-EYFP, p53flox, Nkx3.1CreERT2, and Ptenflox targeted mutations and may be useful when studying prostate cancer.
Corinne Abate-Shen, Columbia University Medical Center
The Nkx3.1CreERT2 targeting vector was designed to insert a cre/ERT2 fusion gene (Cre recombinase fused to a human estrogen receptor ligand binding domain) followed by a polyadenylation sequence into the translation start site of the NK3 homeobox 1 (Nkx3-1) gene. The construct also included a self-excising ACN cassette for positive selection and a PGK-tk (thymidine kinase) cassette for negative selection. The ACN cassette, containing the neomycin resistance gene and Cre recombinase gene under the control of angiotensin-converting enzyme promoter, is flanked by loxP sites. Cre-mediated recombination during spermatogenesis removes the neo cassette leaving one loxP site. The construct was electroporated into 129S6/SvEvTac-derived TC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred with C57BL/6 mice to establish a colony.
Another targeting vector designed to insert a loxP-flanked neo cassette in intron 10, and a third loxP site downstream of exon 11 of the breast cancer 2, early onset (Brca2) gene. The construct was electroporated into 129P2/OlaHsd-derived E14IB10 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a Cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exon 11, intact floxed-neo cassette, or excision of both exon 11 and the neo cassette. Correctly targeted ES cells, containing only the floxed-exon 11, were injected into C57BL/6 blastocysts and resulting chimeric mice were bred with FVB/N mice. Resulting offspring were bred to Nkx3.1CreERT2 mice.
This new strain, Stock No. 033754, was generated by crossing the resulting double allelic offspring to Rosa26-CAG-LSL-EYFP mice (Stock No. 007903), to p53flox mice (Stock No. 008462), and subsequently to Ptenflox mice (Stock No. 006440). Mice are maintained on a mixed genetic background that contains mostly C57BL/6.
Currently there are no related genes or alleles for this strain.
When maintaining a live colony, all alleles may be kept homozygous.
When using the NPp53BR2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #033754 in your Materials and Methods section.