These Nlrp3tm3Hhf (or Nlrp3D301NneoR) knock-in mice serve as a constitutive knock-out of the Nlrp3 gene. This line may be useful in studying the role of cryopyrin in the regulation of autoinflammatory diseases.
Hal M Hoffman, UCSD
Nlrp3D301NneoR mice contain a floxed neomycin cassette in intron 2 of the NLR family, pyrin domain containing 3 gene, Nlrp3, in opposite orientation to gene (neoR), abolishing gene expression. These mice also contain a point mutation in exon 3, which results in a missense mutation, D301N corresponding to human amino acid 303. This mutation is commonly found in humans with neonatal onset multisystem inflammatory disease (NOMID) which is characterized by systemic inflammation and skeletal abnormalities. NLRP3 encodes the protein cryopyrin, which is a cytosolic nucleotide-binding domain and leucine-rich repeat containing (NLR) protein expressed in white blood cells and chondrocytes. Cryopyrin controls the formation of the inflammasome which regulates the immune system's response to injury, toxins, and infection by cleaving interleukin (IL)-1β. NLRP3 mutations are known to cause autoinflammatory diseases known as cryopyrin-associated periodic syndromes (CAPS) such as NOMID, Muckle-Wells syndrome (MWS), and familial cold autoinflammatory syndrome (FCAS). Heterozygotes are viable and fertile. When bred to mice that express Cre recombinase, resulting offspring will have the floxed-neoR deleted in the cre-expressing tissues, allowing expression of the mutated gene.
For example, when bred to mice ubiquitously expressing zona pellucida glycoprotein 3 (Zp3)-cre mice, Nlrp3D301NneoR expression results in systemic inflammation, growth retardation, growth plate disorganization, and low bone mass, and accelerated bone resorption.
A targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette, in reverse orientation to the gene, into intron 2 of the NLR family, pyrin domain containing 3 gene, Nlrp3. A point mutation was introduced into exon 3, resulting in a missense mutation, D301N corresponding to human amino acid 303. This mutation is commonly found in humans with cryopyrin-associated periodic syndromes (CAPS). The construct was electroporated into 129/SvJ-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to C57BL/6NCrl mice for at least 9 generations to establish a colony. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.
|Allele Name||targted mutation 3, Hal M Hoffman|
|Allele Type||Targeted (Conditional ready (e.g. floxed), Humanized sequence, No functional change)|
|Gene Symbol and Name||Nlrp3, NLR family, pyrin domain containing 3|
|Promoter||Nlrp3, NLR family, pyrin domain containing 3, mouse, laboratory|
|Strain of Origin||129|
|Molecular Note||A targeting vector was designed to insert a loxP-flanked neomycin resistance (neo) cassette, in reverse orientation to the gene, into intron 2. A point mutation was introduced into exon 3, corresponding to human amino acid 303, resulting in a missense mutation, D301N, commonly found in humans with cryopyrin-associated periodic syndromes (CAPS).|
|Mutations Made By|| |
Hal Hoffman, UCSD
When maintaining a live colony, homozygous mice may be bred together.
When using the Nlrp3D301NneoR mouse strain in a publication, please cite the originating article(s) and include JAX stock #017971 in your Materials and Methods section.