C57BL/6-Pgbd5^tm1.1Aken/J

Stock number: 037535
Product name: Pgbd5-floxed
Research areas: Cancer Research, Neurobiology Research, Research Tools

Description

Pgbd5-floxed mice possess loxP sites flanking exon 4 of the piggyBac transposable element derived 5 (Pgbd5) gene. These mice may be useful for studying the role of Pgbd5 in neuronal development and plasticity as well as genomic rearrangements and instability in various cancers.

Detailed description

PiggyBac transposable element derived 5 (Pgbd5) is one of the most highly conserved vertebrate transposases, which mediates cut-and-paste DNA transposition of piggyBac transposons. PGBD5 is expressed during embryogenesis and neurogenesis, but also found in increased levels in many solid tumors such as medulloblastomas, neuroblastomas, Ewing sarcomas, desmoplastic small round cell tumors, and rhabdomyosarcomas. Mice that are homozygous for this floxed allele are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 4 deleted in the cre-expressing tissues. These Pgbd5-floxed mice may be useful for studying the role of Pgbd5 in the developing nervous system and tumorigenesis in multiple cancers.

The donating laboratory created a Pgbd5 null allele by crossing these Pgbd5-floxed mice to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J (Stock No. 003724). The homozygous null mice did not express any Pgbd5 exon 4 mRNA in cerebellum as determined by in situ hybridization. Homozygous null mice also did not express measurable PGBD5 protein utilizing immunofluorescence microscopy with rabbit polyclonal antibodies. Unlike wildtype littermates, Pgbd5 null mice demonstrate attention, learning, motor and seizure deficits as early as 30 days of age. Pgbd5 null mice are also resistant to medulloblastoma development.

Development

The Pgbd5-floxed targeting vector was designed to insert an FRT-flanked LacZ and neoR selection cassette upstream of exon 4 of the piggyBac transposable element derived 5 (Pgbd5) gene. The targeting cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene, a second FRT site and a second loxP site. A third loxP site is inserted downstream of exon 4. The construct was electroporated into C57BL/6NTac-derived iTL IC1 embryonic stem (ES) cells and correctly targeted ES cells were injected into BALB/c blastocysts. The resulting chimeric mice were bred with mice expressing Flp recombinase on a C57BL/6 background to remove the neo cassette. Progeny were crossed to C57BL/6J (Stock No. 000664) mice to remove the Flp-expressing transgene. The resulting Pgbd5-floxed mice, with LacZ/neo cassette removed and floxed exon 4 intact, were bred to and maintained on a C57BL/6J genetic background by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Pgbd5^tm1.1Aken
Name: targeted mutation 1.1, Alex Kentsis
MGI accession ID: MGI:7334716
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Pgbd5
Marker name: piggyBac transposable element derived 5
Chromosome: 8
Strain of origin: C57BL/6NTac
Molecular note: The targeting cassette is composed of an FRT site, lacZ sequence, a loxP site, a neomycin resistance gene, a second FRT site, and a second loxP site inserted upstream of exon 4. A third loxP site is inserted downstream of exon 4. FLP-mediated recombination removed the lacZ and neo cassette.