B6(SJL)-Pdcd2^{tm1c(EUCOMM)Wtsi}/RbsnJ

Stock number: 026428
Research areas: Apoptosis Research, Cell Biology Research, Research Tools

Description

Pdcd2^flox mice possess loxP sites flanking exon 2 of the programmed cell death 2 gene. This strain may be useful useful for studying cellular proliferation and cell cycle regulation during embryonic development.

Detailed description

Pdcd2^flox mice possess loxP sites flanking exon 2 of the programmed cell death 2 (Pdcd2) gene. PDCD2 is a zinc finger MYND domain-containing protein that plays a role in the regulation of cellular proliferation, particularly in embryonic stem cells (ESCs), mouse embryonic fibroblast (MEFs) cells, human cancer cells and during early mouse embryonic development. Homozygotes are viable and fertile. When bred to mice that express Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues.

For example, when bred to B6.129-Gt(ROSA)26Sor^{tm1(cre/ERT2)Tyj}/J mice (Stock No. 008463) with widespread expression of tamoxifen inducible cre, and also carrying a Pdcd2^lacZ KO allele, ex vivo cultured ESCs and MEFs display defects in progression from the G1 to the S phase of cell cycle, with increased levels of p53 protein and p53 target genes.

Development

C57BL/6N-A^tm1Brd-derived JM8A3.N1 embryonic stem (ES) cells containing a targeted mutation of the programmed cell death 2 (Pdcd2) gene were obtained from The European Conditional Mouse Mutagenesis Program (EUCOMM). Initially, this Pdcd2 allele had, from 5’ to 3’, a frt site, an En2 splice acceptor cassette containing a β-galactosidase sequence, and loxP-flanked neomycin cassette, containing a frt site, inserted downstream of exon 1. A third loxP site was inserted downstream of exon 2. Correctly targeted ES cells were injected into C57BL/6J blastocysts and resulting chimeric mice from clone EPD0372_1_A09 were bred C57BL/6J mice. Offspring were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J transgenic mice (Stock No. 005703) to delete the splice acceptor cassette and the neo cassette, leaving exon 2 floxed. Progeny were crossed to remove the Flp-expressing transgene. Pdcd2^flox mice were bred to C57BL/6J mice for at least 9 generations. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Pdcd2^{tm1c(EUCOMM)Wtsi}
Name: targeted mutation 1c, Wellcome Trust Sanger Institute
MGI accession ID: MGI:5609169
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Pdcd2
Marker name: programmed cell death 2
Chromosome: 17
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The cassette is composed of from 5' to 3' an frt site, an En2 splice acceptor cassette containing a beta--galactosidase sequence, and loxP-flanked neomycin cassette, containing a frt site, inserted downstream of exon 1. A third loxP site was inserted downstream of exon 2. The critical exon is thus flanked by loxP sites. FLP mediated recombination removed the FRT-flanked lacZ and neo cassette leaving exon 2 flanked by loxP sites. Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml.