NOD.Cg-Capn3^em5Lutzy Prkdc^scid Il2rg^tm1Wjl/VitaJ

Stock number: 038279
Product name: NSG.Capn3 KO
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Mouse/Human Gene Homologs, Neurobiology Research, Research Tools, Virology Research

Description

NSG.Capn3 knock-out is a CRISPR/Cas9 generated mutant of the calpain 3 gene Capn3 gene carrying a 1759 nucleotide deletion on an NSG (Stock No. 005557) background. This severely immunodeficient strain provides an effective model for human cell transplantation/engraftment when studying limb-girdle muscular dystrophy type 2A.

This mouse model was made in conjunction with Coalition to Cure Calpain3 foundation.

Of note, the Capn3^em5Lutzy allele is available on a 129/SvJaeJ genetic background as Stock No. 031211.

Detailed description

NSG.Capn3^em5Lutzy KO mice are NOD.scid.Il2Rg^null ("NSG"; Stock No. 005557) animals also harboring a CRISPR-generated 1759 nucleotide deletion knock-out mutation of the calpain 3 (Capn3)gene.

NSG mice are immunodeficient: they have no mature T cells or B cells, lack functional natural killer (NK) cells, have reduced numbers of lymphocytes and myeloid dendritic cells, and are deficient in cytokine signaling. The NSG platform is permissive for xenograft/human tumor growth. The targeted Capn3 gene encodes a muscle tissue-specific protease. Mutations in this gene have been associated with limb-girdle muscular dystrophy type 2A.

Mice that are homozygous for the Capn3^em5Lutzy allele, homozygous for the Prkdc^scid allele and homozygous or hemizygous for the X-linked Il2rg^tm1Wjl allele are viable and fertile. No significant differences in muscle phenotype were noted in male NSG.Capn3^em5Lutzy KO mice with respect to baseline maximal force production in the tibialis anterior (TA) by 7 months of age. A mild deficit in NSG.Capn3^em5Lutzy KO females was noted in the baseline maximal force production. Following myotoxic injury, NSG.Capn3^em5Lutzy KO recover force mostly at the same rate as age-matched NSG controls with a mild deficit in recovery from injury in males. These mice may be useful for human cell transplantation/engraftment when studying limb-girdle muscular dystrophy type 2A.

Maintaining these severely immunocompromised mice under specific pathogen-free conditions is recommended to promote their overall health. Please see our NSG housing and breeding information for additional details.

This mouse model was made in conjunction with Coalition to Cure Calpain3 foundation.

Development

NSG.Capn3^em5Lutzy KO mice are NOD.scid.Il2Rg^null ("NSG"; Stock No. 005557) animals also harboring a CRISPR-generated 1759 nucleotide deletion knock-out mutation of the calpain 3 (Capn3) gene.

NSG mice are non-obese diabetic animals (NOD/ShiLtJ) harboring the spontaneous severe combined immunodeficiency mutation (Prkdc^scid) on chromosome 16 and the Il2Rg^null mutation at ~101 Mbp on the X chromosome (Il2rg^tm1Wjl created by Dr. Warren J. Leonard [NIH]). NSG mice are described and available from The Jackson Laboratory Repository as Stock No. 005557.

The Capn3^em5Lutzy allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing at The Jackson Laboratory. Plasmids encoding a signal guide RNA designed to introduce a 1759 nucleotide deletion in the calpain 3 (Capn3) gene and the cas9 nuclease were introduced into the cytoplasm 129S4/SvJaeJ-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and maintained on an 129S4/SvJaeJ (Stock No. 009104) background.

On behalf of the Cure for Calpain 3 Coalition, the 129SvJaeJ.Capn3^em5Lutzy (Stock No. 031211) KO mice were backcrossed on to the NSG background for several generations by the donating laboratory. As of June 2024, an additional three generations of breeding to "NSG" mice (Stock No. 005557) was performed at The Jackson Laboratory before intercrossing to homozygosity.

Allele

Symbol: Capn3^em5Lutzy
Name: endonuclease-mediated mutation 5, Cathy Lutz
MGI accession ID: MGI:6270392
Generation method: Endonuclease-mediated
Attributes: Null/Knockout
Marker symbol: Capn3
Marker name: calpain 3
Marker synonyms: CANP3; CANPL3; Capa3; Capa-3; LGMD2; LGMD2A; LGMDD4; LGMDR1; Lp82; Lp84; Lp85; nCL-1; p94
Chromosome: 2
Strain of origin: 129S4/SvJaeJ
Molecular note: CRISPR/Cas9 genome editing is used to introduce a 1759 nucleotide deletion.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Marker synonyms: DNAPDcs; DNAPK; DNA-PK; DNAPKc; DNA-PKC; DNA-PKcs; DNPK1; DOXNPH; dxnph; HYRC; HYRC1; IMD26; p350; S473K; slip; XRCC7
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: [KO]gammac; CD132^-; gammac^-; gc^-; Il2rg^tm1Wjll; IL2Rgamma^null
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Marker synonyms: [g]c; Ab2-183; Cd132; CIDX; common cytokine receptor gamma chain; common gamma chain; gamma C receptor; gamma(c); gammaC; gc; IL-2RG; IMD4; P64; SCIDX; SCIDX1
Chromosome: X
Strain of origin: 129S4/SvJae
Site of expression: Primarily lymphoid cells.
Molecular note: A neomycin resistance cassette replaced part of exon 3 and all of exons 4 - 8 of the gene, resulting in the loss of most of the extracellular domain and all of the transmembrane and cytoplasmic domains of the protein.