C57BL/6-Nfe2l2^tm1.1Sred/SbisJ

Stock number: 025433
Product name: Nrf2^flox
Research areas: Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

Nrf2^flox mice possess loxP sites flanking exon 5 of the Nfe2l2 gene. Exposure to Cre recombinase removes the floxed sequence - creating a null allele. This strain may be useful for generating conditional mutations in applications related to wound repair and inflammation.

Detailed description

The targeted Nfe2l2 gene, also known as Nrf2, encodes a transcription factor that is involved in oxidative stress response. These Nrf2^flox mice possess loxP sites on either side of exon 5 which encodes the DNA binding domain. Mice homozygous for this Nrf2^flox allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 5 deleted in the cre-expressing tissues. Removal of the floxed sequence creates a null allele.

For example, when bred to a strain with Cre recombinase expression in lung Clara cells (Scgb1a1-cre), this mutant mouse strain may be useful in studies of lung injury and inflammation (Reddy et al. 2011).

Furthermore, when crossed to a retinal-specifc Cre expressing strain, such as Six3-cre (Stock No. 019755) or an endothelial-specific Cre such as Cdh5-cre (Stock No. 006137), the deletion of Nrf2 impairs the revascularization of the retina following oxygen-induced retinopathy (OIR) (Wei et al. 2015).

Development

A targeting vector containing a PGK-Neo selection cassette was utilized in the construction of this mutant. A FRT site, a loxP site and this selection cassette followed by a second FRT site and a second loxP site was inserted downstream of exon 5 of the targeted gene. Another loxP site was inserted upstream of exon 5. This construct was electroporated into unspecified C57BL/6-derived embryonic stem (ES) cells which were transiently transfected with a FLPe recombinase vector to remove the selection cassette. ES cells in which Cre recombination resulted in exon 5 being flanked by loxP sites were injected into blastocysts. Resulting chimeric animals were backcrossed to C57BL6/J mice for 10 generations 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 (Stock No. 000664).

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Two of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Nfe2l2^tm1.1Sred
Name: targeted mutation 1.1, Sekhar P Reddy
MGI accession ID: MGI:5463890
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Nfe2l2
Marker name: nuclear factor, erythroid derived 2, like 2
Chromosome: 2
Strain of origin: C57BL/6J
Molecular note: Exon 5 was floxed. Flp-mediated recombination removed the FRT-flanked neo cassette.