These knock-in mice contain a floxed neomycin cassette (neoR) in opposite orientation to a mutated Nlrp3 gene resembling the human mutation associated with Muckle-Wells syndrome. When bred to mice that express Cre recombinase to delete the floxed-neoR, the mutant gene is expressed in cre-expressing tissues of the offspring. These mice may be useful in studying the role of cryopyrin in the regulation of autoinflammatory diseases.
Hal M Hoffman, UCSD
Nlrp3A350VneoR 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). These mice also contain a point mutation in exon 3 which results in a missense mutation, A350V, corresponding to human amino acid 352. This mutation is commonly found in humans with Muckle-Wells syndrome (MWS). 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 MWS, familial cold autoinflammatory syndrome (FCAS), and neonatal onset multisystem inflammatory disease (NOMID). Homozygotes 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 mutant gene.
For example, when bred to B6.129P2-Lyz2tm1(cre)Ifo/J mice (Stock No. 004781), Nlrp3A350VneoR expression in myeloid lineage cells results in postnatal lethality, with 70% of mice dying by 14 days of age. Neutrophilic infiltrates are evident in skin, liver, spleen, joint, sinus, conjunctiva, bone marrow, and tongue. They lack hair and have red scaly skin, and exhibit extreme necrosis of the gut and kidney.
When bred to mice carrying Tg(CAG-(cre/Esr1*)5Amc (Stock No. 004682), tamoxifen-induced Cre-mediated recombination results in hypersecretion of IL1b and an enhanced ability to stimulation T cell differentiation by activated dendritic cells.
When bred to mice carrying Tg(Zp3-cre)3Mrt (Stock No. 003394), Cre recombinase expression in the oocyte results in postnatal lethality from multi-organ failure following inflammation and necrosis.
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, A350V, corresponding to human amino acid 352. 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 chimeric mice were bred to C57BL/6NCrl mice. The donating investigator reports that mutant mice were backcrossed to C57BL/6NCrl for at least ten generations (see SNP note below) prior to sending to The Jackson Laboratory. Upon arrival, heterozygous sperm was frozen. To generate the live colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6NJ inbred females (Stock No. 005304).
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6J genetic background.
|Allele Name||targeted mutation 1, 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|
|Gene Synonym(s)||AGTAVPRL; AII; AVP; C1orf7; CIAS1; CLR1.1; Cias1; Cias1; FCAS; FCAS1; FCU; MWS; Mmig1; NALP3; PYPAF1; Pypaf1; cold autoinflammatory syndrome 1 homolog (human); cryopyrin|
|Promoter||Nlrp3, NLR family, pyrin domain containing 3, mouse, laboratory|
|Strain of Origin||129/Sv|
|Mutations Made By|| |
Hal Hoffman, UCSD
The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided,
their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of
each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders
are needed. Animals typically ship between 10 and 14 weeks from the date of your order. If a second cryorecovery is needed in
order to provide the minimum number of animals, animals will ship within 25 weeks.
The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.
Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.