NOD.129(B6)-Rag1^tm1Mom Cd80^tm1Shr/JbsJ

Stock number: 004673
Product name: NOD.B7-1/Rag1 double knockout
Research areas: Cancer Research, Diabetes and Obesity Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

These Rag1 double knockout mice fail to produce T or B cells.

Detailed description

The NOD.129(B6)Rag1^tm1MomCd80^tm1Shr/JbsJ homozygous mice fail to produce T or B cells. Mice homozygous for both Rag1^tm1Mom and Cd80^tm1Shr are viable and fertile and exhibit no signs of diabetes due to the absence of lymphocytes.

On the NOD background Cd80^tm1Shr alone exacerbates diabetes onset when compared to standard NOD controls. When injected with proteolipid protein in Complete Freund's Adjuvant, NOD mice homozygous for the Cd80^tm1Shr/JbsJ mutation alone develop similar though slightly milder, experimental autoimmune encephalomyelitis (EAE) when compared with NOD controls.

NOD.129(B6)-Rag1^tm1MomCd80^tm1Shr/JbsJ are valuable for unraveling the co-stimulatory pathways of T cell activation as it relates to autoimmune diseases and organ transplantation.

Development

A targeting vector containing a neomycin cassette under the control of the mouse phosphoglycerol kinase (PGK1) promoter was used to disrupt in J1 (129S4) embryonic stem (ES) cells, the second exon of the Cd80 (commonly referenced as B7-1) gene encoding the IgV-like domain. ES cells were injected into C57BL/6J blastocysts. To establish a colony and germ line transmission, chimeric mice were backcrossed to 129S4, prior to intercrossing to generate mice homozygous for Cd80^tm1Shr(Freeman et al, 1993). Garvin et al, 2000, backcrossed Cd80^tm1Shr to NOD for 5 or more generations prior to intercrossing to make a NOD incipient congenic strain homozygous for Cd80^tm1Shr Due to early onset of diabetes and the difficulties in maintaining the strain, Bluestone, et al backcrossed to NOD.129S7(B6)-Rag1^tm1Mom. In 2003,NOD.129(B6)Rag1^tm1MomCd80^tm1Shr/Jbs arrived at The Jackson Laboratory at generation NE9F8. Once rederived into The Jackson Laboratory it was found that markers delineating the Idd13 locus are of 129 origin. NOD.129(B6)Rag1^tm1MomCd80^tm1Shr/JbsJ is maintained homozygous for the targeted mutations of both Cd80 and Rag1.

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag1
Marker name: recombination activating gene 1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

Allele

Symbol: Cd80^tm1Shr
Name: targeted mutation 1, Arlene H Sharpe
MGI accession ID: MGI:1861804
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Cd80
Marker name: CD80 antigen
Chromosome: 16
Strain of origin: 129S4/SvJae
Molecular note: A neomycin selection cassette replaced the exon encoding the IgV-like domain of the protein. RT-PCR analysis on RNA derived from spleen, thymus and LPS-stimulated B cells of homozygous mice demonstrated that no detectable transcript was produced from this allele. FACS analysis on B cells derived from homozygous mice confirmed that no functional protein was encoded by this allele.