NOD.Cg-Prkdc^scid Tg(Ins2-GAD2)2Lt/LtJ

Stock number: 003844
Product name: NOD.scid RIP-GAD A-line
Research areas: Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

These NOD.scid RIP-GAD A-line mice express a transgene containing the human glutamic acid decarboxylase (GAD65) and exhibit a markedly lowered incidence of diabetes.

Detailed description

NOD.Cg-Prkdc^scid Tg(Ins2-GAD2)2Lt/LtJ transgenic mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. Transgenic mice are homozygous for Prkdc^scid and are free of potential insulitic lymphocytes resulting in diabetes resistance. Delayed diabetes onset is observed when splenic lymphocytes from 5 week old pre-diabetic NOD females are adoptively transferred into NOD transgenic mice homozygote for Prkdc^scid when compared to adoptive transfers into control NOD females. Similar to the NOD/ShiLt(Cg)-Tg(Ins2-GAD2)2Lt/J (Stock No. 005870) homozygous transgenic mice are developmentally lethal due to the transgene insertion site (Bridgett et al, Diabetes 1998 47:1848-56).

This model provides a tool for studying the role of GAD2 as an islet autoantigen in the NOD mouse model of Type 1 diabetes.

Development

The transgene Ins2-GAD2 encodes the human glutamic acid decarboxylase2 cDNA under the control of the rat insulin 2 promoter. This transgenic construct was injected into NOD/Lt oocytes and the resulting founders were backcrossed to wildtype siblings. NOD transgenic mice were backcrossed 1 generation to NOD.CB17-Prkdc^scid prior to intercrossing to make homozygote for the Prkdc^scid mutation (Bridgett et al, Diabetes 1998 47:1848-56). In 2005, the Type 1 Diabetes Resource received NOD/LtSz-Prkdc^scid Tg(Ins2-GAD2)2Lt/LtJ at generation N2F31.

Allele

Symbol: Tg(Ins2-GAD2)2Lt
Name: transgene insertion 2, Edward H Leiter
MGI accession ID: MGI:3603349
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(Ins2-GAD2)2Lt
Marker name: transgene insertion 2, Edward H Leiter
Chromosome: 15
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. Densitrometric Southern blot analysis indicates presence of 4-5 copies of the transgene. FISH analysis and pseudo-Giemsa metaphase spread analysis showed integration to have occurred near the centromere of Chr 15. 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.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
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*).