B6-hNLRP3 Mice

Catalog Number: C001616

Strain Name: C57BL/6NCya-Nlrp3tm1(hNLRP3)/Cya

Genetic Background: C57BL/6NCya

Reproduction: Homozygote x Homozygote

 

Strain Description

The Cryopyrin protein, encoded by the NOD-like receptor family pyrin domain-containing 3 (NLRP3) gene, is a core component of the inflammasome in the innate immune system. As a member of the NOD-like receptor (NLR) family, NLRP3 is predominantly expressed in leukocytes and chondrocytes. It participates in the host defense against damage and infection by recognizing pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) to activate immune responses [1]. In its inactive monomeric state, NLRP3 senses intracellular damage signals, such as abnormal protein aggregates and lipid accumulation. Upon activation, NLRP3 oligomerizes, adopting an active conformation and assembling into inflammasome complexes, subsequently activating Caspase-1 to drive the maturation and secretion of pro-inflammatory cytokines, including IL-1β and IL-18 [1-2]. Activated NLRP3 not only induces the release of inflammatory cytokines but also triggers lytic cell pyroptosis. The intracellular components released during pyroptosis can further amplify inflammatory signals, forming a positive feedback loop of autoinflammation. Moreover, IL-1β can exacerbate the inflammatory cascade by stimulating the production of inflammatory markers such as IL-6 and high-sensitivity C-reactive protein (hsCRP) [3-4]. Given NLRP3's upstream position relative to IL-1β/IL-18 and other inflammatory factors, targeting its activity can effectively block the self-reinforcing mechanism of chronic inflammation, providing a significant therapeutic strategy for inflammation-related diseases [5]. The potential therapeutic areas include Alzheimer’s disease, Parkinson’s disease (via neuroinflammation modulation), inflammatory bowel disease, metabolic dysfunction-associated steatohepatitis (MASH), gout, and obesity-related metabolic inflammation [6-7].

B6-hNLRP3 mouse model was generated by replacing the mouse Nlrp3 genomic region (from the ATG start codon to ~500 bp downstream of the 3'UTR) with the human NLRP3 sequence (from 6 bp upstream of the ATG start codon to ~800 bp downstream of the 3'UTR), enabling stable expression of human NLRP3 protein. The B6-hNLRP3 mouse is suitable for studying inflammatory mechanisms, autoimmune diseases, neurodegenerative diseases, and metabolic diseases. It also serves as an ideal tool for human NLRP3-targeted drug development and preclinical efficacy evaluation.

Strain Strategy

Figure 1. Gene editing strategy of B6-hNLRP3 mice. The mouse Nlrp3 locus (ATG start codon to ~500 bp downstream of the 3'UTR) was replaced with the human NLRP3 sequence (6 bp upstream of ATG to ~800 bp downstream of the 3'UTR) via gene editing technology.

 

Applications

  • Studies of the immune system and inflammasome activation mechanisms.
  • Pathological mechanism analysis of cryopyrin-associated periodic syndromes (CAPS).
  • Development, screening, and preclinical pharmacodynamic evaluation of NLRP3-targeted drugs.
  • Neuroinflammation modulation research in neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease).
  • Exploration of mechanisms in metabolic diseases and autoimmune diseases

 

Validation Data

1. Validation of Human NLRP3 and mouse Nlrp3 Gene Expression

Figure 2. RT-qPCR analysis of gene expression in 6-week-old male homozygous B6-hNLRP3 and wild-type (WT) mice. The results showed that homozygous B6-hNLRP3 mice exhibited significant expression of human NLRP3 gene but no mouse Nlrp3 gene expression in the thymus, liver, brain, and spleen. Wild-type mice showed expression of mouse Nlrp3 gene only, with no detectable human NLRP3 gene expression. The relative expression levels of human NLRP3 gene across different tissues were generally consistent with the tissue distribution characteristics of endogenous mouse Nlrp3 (Bars represent mean ± SEM, n=3).

 

2. Validation of Human NLRP3 Protein Expression

Figure 3. Western blot analysis using a human-specific antibody to detect human NLRP3 protein expression in the thymus and lung tissues of 6-week-old wild-type (WT) and B6-hNLRP3 mice. The results demonstrated that B6-hNLRP3 mice successfully expressed human NLRP3 protein. The expression level in lung tissue was higher than that in thymus, which was consistent with the mRNA expression patterns from RT-qPCR analysis.

*Note: The band detected in wild-type mice is due to cross-reactivity of the antibody, as confirmed by RT-qPCR results, B6-hNLRP3 mice specifically express human protein.

 

References
[1]Xu J, Núñez G. The NLRP3 inflammasome: activation and regulation. Trends Biochem Sci. 2023 Apr;48(4):331-344.
[2]Moretti J, Blander JM. Increasing complexity of NLRP3 inflammasome regulation. J Leukoc Biol. 2021 Mar;109(3):561-571.
[3]Sims JE, Smith DE. The IL-1 family: regulators of immunity. Nat Rev Immunol. 2010 Feb;10(2):89-102.
[4]Booshehri LM, Hoffman HM. CAPS and NLRP3. J Clin Immunol. 2019 Apr;39(3):277-286.
[5]Swanson KV, Deng M, Ting JP. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol. 2019 Aug;19(8):477-489.
[6]Yao J, Sterling K, Wang Z, Zhang Y, Song W. The role of inflammasomes in human diseases and their potential as therapeutic targets. Signal Transduct Target Ther. 2024 Jan 5;9(1):10.
[7]Ma Q. Pharmacological Inhibition of the NLRP3 Inflammasome: Structure, Molecular Activation, and Inhibitor-NLRP3 Interaction. Pharmacol Rev. 2023 May;75(3):487-520.