A New Mouse Strain with a Mutation in the NFE2L2 (NRF2) Gene

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Evgeniy S. Egorov¹, Natalia D. Kondratenko^2,3, Olga A. Averina^2,4,5, Oleg A. Permyakov^4,5, Maria A. Emelyanova^4,5, Anastasia S. Prikhodko^1,2, Ludmila A. Zinovkina¹, Petr V. Sergiev^2,4,5, and Roman A. Zinovkin^2,6,a*

¹Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119991 Moscow, Russia

²Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991 Moscow, Russia

³Russian Clinical Research Center for Gerontology, Ministry of Health of the Russian Federation, Pirogov Russian National Research Medical University, 129226 Moscow, Russia

⁴Institute of Functional Genomics, Lomonosov Moscow State University, 119991 Moscow, Russia

⁵Faculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia

⁶HSE University, 101000 Moscow, Russia

^* To whom correspondence should be addressed.

Received September 26, 2023; Revised November 22, 2023; Accepted November 23, 2023

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Transcription factor NRF2 is involved in inflammatory reactions, maintenance of redox balance, metabolism of xenobiotics, and is of particular interest for studying aging. In the present work, the CRISPR/Cas9 genome editing technology was used to generate the NRF2^ΔNeh2 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 NRF2^wt/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 the NRF2^{ΔNeh2/ΔNeh2} homozygotes showed impaired resistance to oxidative stress compared to the wild-type MEFs. The tissues of homozygous NRF2^{ΔNeh2/ΔNeh2} animals had a decreased expression of the NRF2 target genes: NAD(P)H:Quinone oxidoreductase-1 (Nqo1); aldehyde oxidase-1 (Aox1); glutathione-S-transferase A4 (Gsta4); while relative mRNA levels of the monocyte chemoattractant protein 1 (Ccl2), vascular cell adhesion molecule 1 (Vcam1), and chemokine Cxcl8 was increased. Thus, the resulting mutation in the Nfe2l2 gene coding for NRF2, partially impaired function of this transcription factor, expanding our insights into the functional role of the unstructured N-terminus of NRF2. The obtained NRF2^ΔNeh2 mouse line can be used as a model object for studying various pathologies associated with oxidative stress and inflammation.

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KEY WORDS: transcription factor NRF2, transgenic animals, inflammation, oxidative stress

DOI: 10.1134/S0006297923120039

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