| Primary Identifier | MGI:5910172 | Allele Type | Targeted |
| Attribute String | Dominant negative, Humanized sequence, Inserted expressed sequence, Null/knockout | Gene | Nlrp3 |
| Transmission | Germline | Strain of Origin | 129S6/SvEvTac |
| Is Recombinase | false | Is Wild Type | false |
| molecularNote | A Replacer vector containing a mutated loxP site and a mutated human ortholog of the mouse gene was injected into Nlrp3tm1Bhk ES cells. In these ES cells the endogenous locus was replaced with an FRT site flanked neomycin resistance gene cassette which was then removed through flp-mediated recombination. The cells also contain the same mutated loxP site as the Replacer vector. The injected ES cells were subjected to cre-mediated recombination to integrate the vector into the genome. The resulting allele corrects the null allele of the ES cells by inserting the human ortholog into the location of the endogenous gene. The human gene was engineered with a G>A point mutation changing codon 303 from aspartic acid to asparagine (p.Asp303Asn), a variant or polymorphism associated with cryopyrin-associated periodic syndromes (CAPS). The inserted gene is flanked by an upstream mutated loxP site and a downstream FRT site. Expression of the inserted sequence was confirmed by qPCR. |
