FVB.129X1(B6)-Nfe2l2^tm1Ywk/JwkimJ

Stock number: 034618
Product name: FVB.Nrf2^-
Strain synonyms: Nrf2 KO on FVB

Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

Nfe2l2^tm1Ywk (NRF2) knock-out mice on the FVB/NJ background 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.

Of note, The Jackson Laboratory Repository also offers Nrf2^- mice on a C57BL/6 genetic background (Stock No. 017009).

Detailed description

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. The phenotype summarized below is for the parental line: Nrf2^- mice on a C57BL/6 genetic background (Stock No. 017009). It should be noted that the phenotype of this FVB/NJ-congenic Nrf2^- line (Stock No. 034618) could vary from that of the parental line from which it was derived.

In this NRF2 knock-out 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. These mice were subsequently backcrossed to C57BL/6 for at least 10 generations before being donated to The Jackson Laboratory in 2011 as Stock No. 017009.

In 2016, Dr. Jung-Whan Kim purchased some Stock No. 017009 animals (~N12 on C57BL/6) and then backcrossed them at least 12 generations to FVB/NJ inbred mice (from Stock No. 001800). The resulting FVB/NJ-congenic strain is called FVB.Nrf2^-. Of note, the donating investigator reports that, at least once during backcrossing, a Nrf2 KO female was bred to an FVB/NJ inbred male (thus the Y chromosome of the congenic strain is of FVB/NJ origin). In 2020, FVB.Nrf2^- males (white coat color) were sent to The Jackson Laboratory Repository as Stock No. 034618. Upon arrival, males were used to cryopreserve sperm. To establish our live colony, an aliquot of frozen sperm was used to fertilize FVB/NJ oocytes.

Allele

Symbol: Nfe2l2^tm1Ywk
Name: targeted mutation 1, Yuet Wai Kan
MGI accession ID: MGI:2385926
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: Nrf2-; Nrf2 KO
Marker symbol: Nfe2l2
Marker name: nuclear factor, erythroid derived 2, like 2
Marker synonyms: IMDDHH; Nrf2; Nrf-2
Chromosome: 2
Strain of origin: 129X1/SvJ
Molecular note: A 4.2 kb segment, including 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. Western blot analysis confirmed the absence of protein expression in liver samples from homozygous mutant mice.