B6;129S-Pdyn^{tm1.1(cre)Mjkr}/LowlJ

Stock number: 027958
Product name: Pdyn-IRES-Cre
Research areas: Neurobiology Research, Research Tools

Description

Pdyn-IRES-Cre mice express Cre recombinase under the direction of the Pdyn (prodynorphin) promoter in the paraventricular hypothalamus making them useful for cell-specific neuron mapping.

Detailed description

Pdyn-IRES-Cre mice have an internal ribosomal entry sequence (IRES) fused to Cre recombinase inserted downstream of the endogenous Pdyn (prodynorphin) gene. Prodynorphin is a precursor to multiple active peptides that are released after processing by proprotein convertase 2 (PC2). This cleaving occurs within synaptic vesicles in the presynaptic terminal of the paraventricular hypothalamus and results in the production of the opioid receptors dynorphin A, dynorphin B and α/β-neo-endorphin. Homozygous mice are viable and fertile. When these mice are bred with mice containing loxP-flanked sequence, Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing neurons of the offspring.

Development

A targeting construct was designed to insert an internal ribosomal entry sequence (IRES) fused to Cre recombinase, and a frt-flanked neomycin resistance (neo) cassette, downstream of the stop codon of the Pdyn (prodynorphin). This construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric mice were bred to 129S4/SvJaeSor-Gt(ROSA)26Sor^tm1(FLP1)Dym/J transgenic mice (Stock No. 003946) to remove the neo cassette. These Pdyn-IRES-Cre mice were maintained on a mixed background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Pdyn^{tm1.1(cre)Mjkr}
Name: targeted mutation 1.1, Michael J Krashes
MGI accession ID: MGI:5562873
Generation method: Targeted
Attributes: Recombinase-expressing
Marker symbol: Pdyn
Marker name: prodynorphin
Chromosome: 2
Strain of origin: 129S6/SvEvTac
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Cre recombinase is expressed under the direction of the Pdyn promoter in the paraventricular hypothalamus.
Molecular note: Using recombineering methods, a selection cassette containing an internal ribosomal entry sequence (IRES) linked to cre and an frt-flanked kanamycin gene was inserted immediately downstream of the stop codon in exon 2 of the Pdyn gene in a BAC. This construct was used to isolate a targeting plasmid with the cre-kanamycin cassette with upstream and downstream Pdyn homology arms. ES cells were targeted and used for blastocyst injection. Chimeras were bred for germline transmission. The selection marker was excised by breeding to Flp deleter animals.