A new mouse strain with mutation in the NFE2L2 (NRF2 gene

Abstract

The transcription factor NRF2 is involved in inflammatory reactions, maintenance of redox balance, metabolism of xenobiotics, and is of particular interest for aging studies. In the present work, CRISPR/Cas9 genome editing technology was used to generate NRF2ΔNeh2 mice containing a substitution of eight amino acid residues at the N-terminus of the NRF2 protein, upstream of the functional Neh2 domain, which ensures binding of NRF2 to its inhibitor KEAP1. Heterozygote NRF2wt/ΔNeh2 mice gave birth to homozygous mice with lower than expected frequency, accompanied by their increased embryonic lethality and visual signs of anemia. Mouse embryonic fibroblasts (MEFs) from NRF2ΔNeh2/ΔNeh2 homozygotes showed impaired resistance to oxidative stress compared to wild-type MEFs. The tissues of homozygous NRF2ΔNeh2/ΔNeh2 animals had a decreased expression of NRF2 target genes: NAD(P)H: Quinone oxidoreductase-1 (Nqo1); aldehyde oxidase-1 (Aox1); glutathione-S-transferase A4 (Gsta4); while the relative mRNA level of monocyte chemoattractant protein 1 (Ccl2), vascular cell adhesion molecule 1 (Vcam1) and chemokine Cxcl8 was increased. Thus, the resulting mutation in the Nfe2l2 gene partially impaired the function of this transcription factor, expanding the insights into the functional role of the unstructured N-terminus of NRF2. The obtained NRF2ΔNeh2 mouse line can be used as a model object for studying various pathologies associated with oxidative stress and inflammation.