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

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Abstract

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 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 the NRF2ΔNeh2/ΔNeh2 homozygotes showed impaired resistance to oxidative stress compared to the wild-type MEFs. The tissues of homozygous NRF2ΔNeh2/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 NRF2ΔNeh2 mouse line can be used as a model object for studying various pathologies associated with oxidative stress and inflammation.

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Abbreviations

AOX1:

aldehyde oxidase-1

ARE:

antioxidant response element

CCL2:

monocyte attractant protein

GOx:

glucose oxidase

GSTA4:

glutathione-S-transferase A4

HMOX1:

hemoxygenase-1

KEAP1:

Kelch-like ECH-associated protein 1

MEFs:

mouse embryonic fibroblasts

Neh:

Nrf2-ECH homology

NQO1:

NAD(P)H: quinone oxidoreductase

NRF2:

nuclear factor 2 related to erythroid factor 2

ROS:

reactive oxygen species

VCAM-1:

vascular cell adhesion molecule 1

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Acknowledgments

The authors would like to thank Olga Yurievna Pletjushkina for her invaluable help in working with the MEFs.

Funding

This work was financially supported by the Russian Science Foundation (project no. 21-64-00006).

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Authors and Affiliations

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

    Evgeniy S. Egorov, Anastasia S. Prikhodko & Ludmila A. Zinovkina

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

    Natalia D. Kondratenko, Olga A. Averina, Anastasia S. Prikhodko, Petr V. Sergiev & Roman A. Zinovkin

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

    Natalia D. Kondratenko

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

    Olga A. Averina, Oleg A. Permyakov, Maria A. Emelyanova & Petr V. Sergiev

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

    Olga A. Averina, Oleg A. Permyakov, Maria A. Emelyanova & Petr V. Sergiev

  6. HSE University, 101000, Moscow, Russia

    Roman A. Zinovkin

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  1. Evgeniy S. Egorov
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Contributions

E.S.E. conducting experiments, writing the text, N.D.K. conducting experiments, statistical analysis, and preparation of figures (E.S.E. and N.D.K. contributed equally to the work), O.A.A. working with laboratory animals, conducting experiments, discussing the results of the study, O.A.P., M.A.E., and A.S.P. conducting experiments, L.A.Z. and P.V.S. conducting experiments, discussing the results of the study, R.A.Z. writing the text, concept and guiding the work.

Corresponding author

Correspondence to Roman A. Zinovkin.

Ethics declarations

The work with mice was approved by the local bioethics committee “Institute of Mitoengineering MSU” LLC, protocol #79 of April 28, 2015. The authors of this work declare that they have no conflicts of interest.

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Egorov, E.S., Kondratenko, N.D., Averina, O.A. et al. A New Mouse Strain with a Mutation in the NFE2L2 (NRF2) Gene. Biochemistry Moscow 88, 1987–1996 (2023). https://doi.org/10.1134/S0006297923120039

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