B6.129-Nox4^tm1Kkr/J

Stock number: 022996
Product name: NOX4 KO
Research areas: Internal/Organ Research

Description

Kidneys in these Nox4 knockout mice exhibit increased oxidative stress. They are suitable for use in applications related to the study of oxidative stress-induced tissue damage in disease models for idiopathic pulmonary fibrosis, liver fibrosis, diabetic nephropathy and renal obstruction.

Detailed description

A neo cassette replaces exon 4 of the of the NADPH oxidase 4 (Nox4) gene, abolishing gene expression in these knock-out mice. NOX4 is widely expressed in many cell types and generates reactive oxygen species (ROS). NOX4 has been implicated in the development of fibrotic diseases, in particular idiopathic pulmonary fibrosis (IPF), liver fibrosis, and diabetic nephropathy. These KO mice do not express NOX4 in any tissues including lung, kidney, and spleen. Mice are generally healthy and do not display a notable phenotype with the exception of a moderate tendency for accelerated increase in weight. Homozygous mice are viable and fertile. When used in a model of chronic renal injury by obstruction, mutant mice exhibit interstitial fibrosis and tubular apoptosis, with lower interstitial capillary density and reduced expression of hypoxia-inducible factor-1α and vascular endothelial growth factor in obstructed kidneys. NOX4-deficient kidneys exhibit increased oxidative stress.

Development

A targeting vector was designed to replace exon 4 of the NADPH oxidase 4 (Nox4) gene with a neomycin resistance (neo) cassette. The construct was electroporated into 129Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6J females. These mice were backcrossed 5 generations to C57BL/6J mice using a marker-assisted, speed congenic approach (see SNP note below). Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 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. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 of 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/6J ; C57BL/6N genetic background.

Allele

Symbol: Nox4^tm1Kkr
Name: targeted mutation 1, Karl-Heinz Krause
MGI accession ID: MGI:5502326
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Nox4
Marker name: NADPH oxidase 4
Chromosome: 7
Strain of origin: 129
Molecular note: A targeting vector was designed to replace exon 4 with a neomycin resistance (neo) cassette. Western blot analysis confirmed the absence of protein in the lung.