NOD/ShiLt-Tg(Ins2-GAD2)1Lt/LtJ

Stock number: 003074
Product name: NOD.RipGAD2- Y line
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research

Description

These NOD.RipGAD2- Y line mice express a transgene containing a human B-cell autoantigen, GAD2, under control of a rat insulin promoter, Ins2.

Detailed description

Transgenic mice are characterized by pancreatic beta cells that express a rat insulin promoter 7, Ins2, regulated transgene encoding a human B-cell autoantigen, GAD2. These mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. This NOD/Lt transgenic demonstrates a Y Chr. integration-site as only male progeny are transgenic. Expression of the transgene in the islets has been confirmed by RT-PCR and Western blot analysis. Southern analysis confirms 17 copies of the transgene are present, compared to the A-line that has 4-5 copies (Stock No. 5870). Elevated levels of GAD2 expression in transgenic B-cells do not influence non-fasting plasma levels of insulin and glucagon. At 50 weeks of age 55% of the transgenic males were diabetic, this is not significantly different from NOD/Lt male cohorts. Insulitis in 9 week old females is not significantly different between transgenics and controls.

This model provides a tool for studying at the role of autoantigens, specifically GAD2, in diabetes.

Development

Glutamic acid decarboxylase 2, GAD2 (commonly referred to as GAD65) and located on Chr. 2, is highly expressed in human and rat beta cells, but little to no expression in mouse beta cells. Because GAD2 is a major candidate autoantigen in human T1D, a human GAD2 transgene under control of the Ins2 promoter was injected directly into NOD/Lt zygotes. In 2006, the T1DR received this transgenic strain at generation F?+33.

Allele

Symbol: Tg(Ins2-GAD2)1Lt
Name: transgene insertion 1, Edward H Leiter
MGI accession ID: MGI:3606176
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: TgN(RIP7hGAD65)Lt4 Y-line; RIP7-hGAD65
Marker symbol: Tg(Ins2-GAD2)1Lt
Marker name: transgene insertion 1, Edward H Leiter
Chromosome: Y
Strain of origin: NOD/ShiLt
Expressed genes: GAD2,glutamate decarboxylase 2,human
Molecular note: The transgene comprises the cDNA sequence encoding human glutamic acid decarboxylase 2 under control of a modified rat insulin 2 promoter composed of 9.5 kb of genomic DNA from upstream of the transcription initiation site fused to the first intron of Ins2. Transmission from the founder male only to male progeny demonstrated integration into the Y chromosome. Densitrometric Southern blot analysis indicates presence of approximately 17 copies of the transgene. Expression in the pancreatic islets of transgenic mRNA has been demonstrated by reverse transcription followed by PCR (RT-PCR), and of GAD2 protein by western blot analysis.