NOD-Tg(Igh-6/Igh-V281)3Jwt/JwtJ

Stock number: 005020
Product name: NOD.IgIns-281H
Research areas: Diabetes and Obesity Research, Research Tools

Description

These NOD.IgIns-281H mice express a transgene containing the variable region of the heavy chain B-cell receptor (VH281) that lacks the high affinity specificity for insulin. The mice develop insulitis but are protected from type I diabetes.

Detailed description

The double transgenic produced by crossing this stock with Stock No. 005018 serves as a control to the double transgenic which results from crossing Stock No. 005018 with Stock No. 005019.

Development

Thomas and Hulbert received the BALB/c IgG1 mouse anti-human insulin monoclonal antibody, mAb125 from the American Type Culture Collection (ATCC). This mab cross-reacts to mouse insulin. It was found that the variable region of the heavy chain of mAb125 has some sequence differences from the germ line sequence that generated this antibody sequence, referred to as V_H-IX-281. When residues 49 and 56 of the variable region of the heavy chain of mAb125 are mutated back to the residues found in the germ line sequence, the resulting antibody no longer binds insulin with high affinity. The construct for Tg(Igh-6/Igh-V281)3Jwt has these germ line reversions (from G to D at residue 49 and from E to V at residue 56). This VDJ sequence, with these two residue changes, was cloned in place of the endogenous variable sequence downstream of the Ighm sequence in the construct previously used by Goodenow et al., to generate anti-hen egg lysozyme transgenes. This new construct encodes an IgM^a constant region with a variable region nearly identical to that of mAB125 but lacking the high affinity specificity for insulin. This construct was microinjected into NOD/Tac oocytes and the transgenic strain has been mated to NOD/Tac for 7 generations. In 2004, N7 mice were backcrossed to NOD/LtJ at The Jackson Laboratory, and are maintained hemizygote x wild type.

Allele

Symbol: Tg(Igh-6/Igh-V281)3Jwt
Name: transgene insertion 3, James W Thomas
MGI accession ID: MGI:3056939
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Igh-6/Igh-V281)3Jwt
Marker name: transgene insertion 3, James W Thomas
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: Ighm,immunoglobulin heavy constant mu,mouse, laboratory; Igh-V,immunoglobulin heavy chain variable region,mouse, laboratory
Molecular note: The construct for Tg(Igh-6/Igh-V281)3Jwt has a germ line reversion encoding D at residue 49 (from G)and another encoding V (from E) at residue 56. The transgene encodes an IgM^a constant region with a variable region nearly identical to that of the BALB/c IgG1 mouse anti-human insulin monoclonal antibody, mAb125, (from the American Type Culture Collection) but lacking the high affinity specificity for insulin.