B6.129X1-Nfe2l2^tm1Ywk/J

Stock number: 017009
Product name: Nrf2^-
Strain synonyms: Nrf2 KO, NRF2-
Research areas: Cancer Research, Cardiovascular Research, Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Mouse/Human Gene Homologs, Neurobiology Research, Research Tools, Sensorineural Research

Description

Nfe2l2^tm1Ywk (NRF2) knockout mice may be useful for studying oxidative stress in the pathogenesis of age-related macular degeneration, diabetes, Parkinson's disease, and other inflammatory degenerative diseases as well as studies of cancer, multiple sclerosis, liver cirrhosis, atherosclerosis, injury and wound healing, and more.

Detailed description

In this NRF2 knockout strain, a β-galactosidase (lacZ) reporter followed by a neomycin resistance (neo) cassette replaces exon 5 and part of exon 4 encoding the nuclear factor, erythroid derived 2, like 2 gene (Nfe2l2) gene, abolishing gene function. Homozygotes are viable, fertile, and normal in size. NRF2, a member of the "cap 'n' collar" (CNC) subfamily of the basic region-leucine zipper transcription factors, is a regulator of endogenous antioxidant protection, microglial function, and chronic neuroinflammation. Inactivation of the CNC, DNA binding, and leucine zipper domains in Nrf2^-/- mice results in age-related macular degeneration (AMD)-like retinal pathology, spontaneous choroidal neovascularization (CNV), increased sensitivity to toxins, impaired adipogenesis, abnormal mitochondria, and an increase in proinflammatory gene expression in microglia and astrocytes. The donating investigator reports that lacZ failed to generate any detectable β-galactosidase activity.

Development

A targeting vector was designed to replace exon 5 and part of exon 4 encoding the nuclear factor, erythroid derived 2, like 2 (Nfe2l2) gene with a β-galactosidase (lacZ) reporter followed by a neomycin resistance (neo) cassette and a polyadenylation sequence. The construct was electroporated into 129X1/SvJ-derived JM-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric mice were bred to C57BL/6 mice to generate a colony of Nrf2^-/- mice. The donating investigator reported that these mice were backcrossed to C57BL/6J (see SNP note below) for at least 10 generations . Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation.

In 2018, 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. One of the 5 markers that determine C57BL/6J from C57BL/6N was found to be segregating. This marker is the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Nfe2l2^tm1Ywk
Name: targeted mutation 1, Yuet Wai Kan
MGI accession ID: MGI:2385926
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Nfe2l2
Marker name: nuclear factor, erythroid derived 2, like 2
Chromosome: 2
Strain of origin: 129X1/SvJ
Molecular note: A 4.2 kb segment, includind part of exon 4 and all of exon 5, was replaced by an in-frame lacZ gene followed by a neomycin selection cassette. Northern blot analysis on RNA derived from various tissues confirmed that a fusion transcript was detectable in intestine of homozygous mice, but no wild-type transcript was detectable in any tissue. However, no fusion protein was detected by histochemical staining in any adult tissue.