B6.129S(C)-Ndufaf2^tm1.1Itl/J

Stock number: 027919
Product name: Ndufaf2 KO
Research areas: Neurobiology Research

Description

Ndufaf2 KO mice may be useful for studying the role of Complex-1 in the onset of mitochondrial encephalopathies.

Detailed description

Ndufaf2 KO mice lack endogenous exon 2 of the NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, assembly factor 2 (Ndufaf2) gene, abolishing gene expression. NDUFAF2 is a molecular chaperone for mitochondrial complex I assembly and is involved in the translocation of protons across the inner mitochondrial membrane during the first step of the mitochondrial respiratory chain. Mutations in NDUFAF2 have been associated with a complex encephalopathy in humans, and mitochondrial dysfunction has been more broadly associated with neurodegenerative diseases such as idiopathic Parkinson's disease, Alzheimer's disease, Huntington's disease and Amyotrophic lateral sclerosis. Mice that are homozygous for this allele are viable and fertile. Cells cultured from these mice have reduced Complex-1 activity and display significant increases in oxidative stress and mitochondrial DNA deletion.

Development

A targeting vector was designed by Dr Matthew LaVoie (Brigham and Women's Hospital and Harvard Medical School) to insert a Lox66 site and a Lox71 site, in opposite orientation, flanking a STOP codon, in reverse orientation, downstream of exon 1 of the NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, assembly factor 2 (Ndufaf2) gene. Also, frt-sites flank a neomycin resistance cassette, and Lox2272 sites surround the neo cassette and exon 2 downstream of the reverse STOP. The construct was electroporated into 129SvEv embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred with B6.C-Tg(CMV-cre)1Cgn/J transgenic mice (Stock No. 006054) to delete the neo cassette and exon 2, while also inverting the STOP codon into a sense orientation. Resulting Ndufaf2 KO mice were bred to C57BL/6J mice for 9 generations to establish a colony. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Ndufaf2^tm1.1Itl
Name: targeted mutation 1.1, inGenious Targeting Laboratory
MGI accession ID: MGI:5521199
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Ndufaf2
Marker name: NADH:ubiquinone oxidoreductase complex assembly factor 2
Marker synonyms: 1810058I14Rik; B17.2L; MC1DN10; mimitin; MMTN; Ndufa12l; RGD1560158
Chromosome: 13
Strain of origin: 129S/SvEv
Molecular note: A targeting vector was designed to insert loxP sites around exon 2 and a neo cassette while inserting an inverted exon 2 with multiple stop codons downstream of the floxed sequence. Cre-mediated recombination removed the exon 2 and neo cassette resulting in the placement of the mutant exon 2 in-frame with the remainder of the Ndufaf2 gene. Western blot analysis confirmed the absence of protein expression in skin fibroblasts.