B6.129P2(FVB)-Igh^tm1.1Jwt/J

Stock number: 029288
Product name: VH125^SD
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research

Description

VH125^SD mice express an anti-insulin VDJ_H-125 site directed to the native IgH locus making them useful for studying peripheral tolerance in autoreactive B lymphocytes.

Detailed description

VH125^SD mice express an anti-insulin VDJ_H-125 site directed to the native IgH locus. Anti-insulin B cells formed in these mice are capable of class-switch recombination, permitting assessment of the IgG class of insulin Autoantibodies. Anti-insulin B cells fail to produce IgG antibodies following T cell-dependent immunization with heterologous insulin, and they exhibit markedly impaired proliferation to anti-CD40 plus insulin in vitro. Costimulation with LPS plus insulin results in anti-insulin B cell proliferation. These VH125^SD mice produce both IgM and IgG2a anti-insulin antibodies following immunization with insulin conjugated to type 1 T cell-independent Brucella abortus ring test antigen (BRT).

Of note, this allele is also available on an NOD congenic background (Stock No. 029397).

Development

The VH125^SD construct contains an anti-insulin VDJ_H-125 site directed to the native IgH locus and a loxP-flanked neomycin resistance cassette in reverse orientation to the gene. The construct was electroporated into 129/Ola embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred with EIIA-Cre mice on a B6 background to remove the neo cassette. Resulting offspring were bred to C57BL/6J mice for at least 15 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Igh^tm1.1Jwt
Name: targeted mutation 1.1, John Williams
MGI accession ID: MGI:5780066
Generation method: Targeted
Attributes: Inserted expressed sequence
Synonyms: VH125^SD
Marker symbol: Igh
Marker name: immunoglobulin heavy chain complex
Chromosome: 12
Strain of origin: 129P2/OlaHsd
Molecular note: The targeting construct replaces the IgJH loci with an anti-insulin rearranged VDJ-H-125 site. A loxP-flanked neomycin resistance cassette, in reverse orientation to the gene, is located upstream. Cre-mediated recombination removed the upstream floxed neo cassette.