NOD/ShiLtJ-Ptpn22^em1Shrm/J

Stock number: 036948
Product name: Ptpn22^R619W
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research

Description

Ptpn22^R619W knockin mice express an arginine to tryptophan amino acid substitution, which is orthologous to human PTPN22^R620W, on the autoimmune type 1 diabetes (T1D) susceptible NOD/LtShiJ background. The mutation enhances onset and penetrance of T1D.

Of note: the Ptpn22^R619Wmutation also is available on the C57BL/6 background, Stock No. 036949.

Detailed description

Ptpn22 (protein tyrosine phosphatase, non-receptor type 22 (lymphoid)) encodes a lymphoid specific intracellular phosphatase that functions in the regulation of T-cell, B-cell and myeloid cell signaling during T cell development and differentiation. Ptpn22^R619W knockin mice were created using CRISPR/cas9 genome editing to express the R619W (arginine to tryptophan) amino acid substitution. R619W is orthologous to human R620W. The mutation is located in the P1 domain and causes diminished binding to the inhibitor tyrosine kinase CSK. The R620W mutation is associated with multiple autoimmune diseases, in particular, type diabetes (T1D). Mice homozygous for the mutation are viable and fertile. Female homozygotes (WW) exhibit increased serum insulin autoantibodies by 4 weeks of age and develop diabetes starting around 5-6 weeks of age; 100% will become diabetic before the age of 21 weeks. The males do not demonstrate this accelerated disease course and are similar to NOD/ShiLtJ males.

Of note: the Ptpn22^R619Wmutation also is available on the C57BL/6 background, Stock No. 036949.

Development

CRISPR/cas9 genome editing was used to create an amino acid substitution at position 619 (arginine to tryptophan, CGG to TGG, R619W) in exon 14 and a silent mutation (CTC to CTT) at amino acid position 616 (creating a BspE1 restriction site) in the Ptpn22 locus on Chromosome 3. Single guide RNA and Cas9 endonuclease were introduced into NOD/ShiLtJ-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to NOD/ShiLtJ for germline transmission. Correctly targeted pups were identified by sequencing and PCR. Mice were bred to NOD/ShiLtJ for at least 2 generations reducing the potential for off-target effects. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize NOD/ShiLtJ (Stock No. 001976) oocytes.

Allele

Symbol: Ptpn22^em1Shrm
Name: endonuclease-mediated mutation 1, Linda A Sherman
MGI accession ID: MGI:6149963
Generation method: Endonuclease-mediated
Attributes: Not Specified
Synonyms: Ptpn22^R619W
Marker symbol: Ptpn22
Marker name: protein tyrosine phosphatase, non-receptor type 22 (lymphoid)
Marker synonyms: 70zpep; LYP; LYP1; LYP2; PEP; PTPN22.5; PTPN22.6; PTPN8
Chromosome: 3
Strain of origin: NOD/ShiLtJ
Molecular note: CRISPR-targeting generated a 4 nucleotide substitution in exon 14 that results in the amino acid substitution of tryptophan for arginine at position 619 (R619W). An additional silent mutation (CTC to CTT) was introduced at amino acid position 616.