NOD.129S4(B6)-Art2a^tm1Fkn Art2b^tm1Fkn/LtJ

Stock number: 008252
Product name: NOD.Art2
Research areas: Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

This Art2a and Art2b targeted mutant strain becomes diabetic at a similar onset and rate as NOD/ShiLt mice. The absence of ART2 expression on T cells from thymus and lymph nodes was confirmed by RT-PCR and FACS analysis. Cd4⁺ and CD8⁺ T cells from ART2 deficient NOD mice are completely resistant to NAD induced apoptosis at all concentrations in vitro. This ART2 deficient stock provides a tool to further dissect the interplay of ADP rybosylation and NAD-catabolizing enzymes and their effect on Tregs and iNKT involved in autoimmune diseases specifically, Type 1 Diabetes.

Detailed description

Art2a, Art2b, commonly referred to as ART2, deficient mice are viable and fertile and do not show any overt phenotype. RT-PCR and FACS analysis confirm the absence of ART2 expression on the cell surface of thymic and lymph node purified T cells. No detectable histological abnormalities were observed in the thymus, spleens, lymph nodes, Peyer's patches, thyroid, brain, lung, kidney, intestine, liver, pancreas and thyroid. The disrupted introgression of ART2 onto the NOD background resulted in no significant difference in diabetes onset or incidence when compared to NOD. When the Cd38 targeted mutation is introduced to the NOD.ART2 deficient stock, the NOD.CD38, ART2 deficient males and females are completely resistant to diabetes. This is a surprising result as the NOD.CD38 deficient stock experiences accelerated diabetes onset and incidence in both males and females. Cd4⁺ and CD8⁺ T cells from ART2 deficient NOD mice are completely resistant to NAD induced apoptosis at all concentrations in vitro.

The ART2 targeted mutation provides a tool to further dissect the interplay of ADP rybosylation and NAD-catabolizing enzymes and their effect on Tregs and iNKT involved in autoimmune diseases specifically, Type 1 Diabetes.

Development

Art2, ADP-ribosyltransferase 2, is a NAD-metabolizing ectoenzyme expressed on the surface of peripheral T cells and shed upon activation. The Art2a and Art2b genes were disrupted via homologous recombination in 129S4/SvJae-derived, J1 ES cells. Most of exon 3, the major coding exon, of Art2a was replaced with a neomycin resistance cassette. The ES cell clone bearing the Art2a targeted mutation was then transfected with a construct which replaced most of the major coding exon of Art2b with a hygromycin resistance cassette. The resulting ES cell clone with two targeted mutations was injected into blastocysts. The resulting chimeric founder's offspring were bred to C57BL/6J prior to 11 generations of crossing to NOD/ShiLt and intercrossing. In 2008, the T1DR received mice homozygous for the Art2a/Art2b mutation at generation N11F16.

Allele

Symbol: Art2b^tm1Fkn
Name: targeted mutation 1, Friedrich Koch-Nolte
MGI accession ID: MGI:2388827
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Art2b
Marker name: ADP-ribosyltransferase 2b
Chromosome: 7
Strain of origin: 129S4/SvJae
Molecular note: The coding region was disrupted by a hygromycin selection cassette inserted by homologous recombination. This construct was used to transfect mice in which the Art2a^tm1Fkn allele had already been created Transcript was undetected in homozygous mutant mice by RT-PCR analysis of T cells isolated from lymph nodes. FACS analysis of thymocytes and lymphocytes indicated a lack of protein expression on the cell surface.
General note: The targeting vector was electroporated into J1 ES cells carrying Art2a^tm1Fkn.

Allele

Symbol: Art2a^tm1Fkn
Name: targeted mutation 1, Friedrich Koch-Noltet
MGI accession ID: MGI:2388822
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Art2a
Marker name: ADP-ribosyltransferase 2a
Chromosome: 7
Strain of origin: 129S4/SvJae
Molecular note: The coding region was disrupted by a neomycin selection cassette inserted by homologous recombination. Transcript was undetected in homozygous mutant mice by RT-PCR analysis of T cells isolated from lymph nodes.
General note: Successfully targeted ES cells were subsequently targeted with the construct for the Art2b allele.