B6.129X1-Dgkz^tm1Gak/J

Stock number: 018861
Research areas: Immunology, Inflammation and Autoimmunity Research

Description

These Dgkz knockout mice exhibit a 5-10% decrease of CD4⁺ and CD8⁺ T cells and an increase of B220⁺ B cells in spleen and lymph nodes, with normal CD4⁺ T cell numbers in the thymus. They may be useful for studying the role of diacylglycerol in T cell development and function.

Detailed description

An internal ribosome entry site (IRES)-GFP followed by a loxP-flanked neomycin resistance cassette replaces the C1 and kinase domains of the diacylglycerol kinase zeta (Dgkz) gene, abolishing gene function. DGKζ is an enzyme expressed in T cells, B cells, mast cells, neutrophils, macrophages, and dendritic cells as well as many other non-hematopoietic cells. DGKζ converts diacylglycerol, a compound essential to normal T cell homeostasis, to phosphatidic acid which has been found to inhibit TCR signaling. These mice have a 5-10% decrease of CD4⁺ and CD8⁺ T cells and an increase of B220⁺ B cells in spleen and lymph nodes, with normal CD4⁺ T cell numbers in the thymus. T cells of DGKζ-deficient mice are hyper-responsive to TCR stimulation, and show enhanced activation of the Ras-ERK cascade and increased expression of activation markers. These mice exhibit an increase in total splenic CD8+ cells in response to viral infection. Homozygotes are viable and fertile.

Development

A targeting vector was designed to replace exons encoding the C1 and kinase domains of the diacylglycerol kinase zeta (Tsc22d1) gene with an internal ribosome entry site (IRES)-GFP followed by a loxP-flanked neomycin resistance cassette. The construct was electroporated into 129X1/SvJ-derived RW4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and the resulting chimeric mice were bred to C57BL/6J mice to generate the mutant colony. These mutant mice were bred to C57BL/6J mice for at least 9 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Dgkz^tm1Gak
Name: targeted mutation 1, Gary A Koretzky
MGI accession ID: MGI:2674915
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Dgkz
Marker name: diacylglycerol kinase zeta
Chromosome: 2
Strain of origin: 129X1/SvJ
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: The diacylglycerol kinase zeta (Dgkz) gene is normally expressed in T cells, B cells, mast cells, neutrophils, macrophages, and dendritic cells as well as many other non-hematopoietic cells.
Molecular note: A cassette containing floxed neo and IRES-GFP replaced 6.1 kb of genomic sequence encompassing exons encoding the C1 and kinase domains. Normal protein was undetected by Western blot analysis of homozygous mutant thymocytes, splencytes, and bone marrow-derived macrophages.