Overview

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.

Donating Investigator

Gary Koretzky, University of Pennsylvania

Genetic overview

Genetic Background	Generation

Dgkz^tm1Gak

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter, Null/Knockout)	Dgkz	diacylglycerol kinase zeta

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Research Applications

Immunology, Inflammation and Autoimmunity Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

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.

Expression Data

Expressed Gene	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.

Control Suggestions

Wild-type from the colony

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Zhong XP; Hainey EA; Olenchock BA; Jordan MS; Maltzman JS; Nichols KE; Shen H; Koretzky GA. 2003. Enhanced T cell responses due to diacylglycerol kinase zeta deficiency. Nat Immunol 4(9):882-90PubMed: 12883552 MGI: J:85149

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Genetics

Dgkz^tm1Gak

Allele Symbol: Dgkz^tm1Gak

Allele Name	targeted mutation 1, Gary A Koretzky
Allele Type	Targeted (Reporter, Null/Knockout)
Allele Synonym(s)	Dgkz-; Dgkzeta-; Dgkzeta KO
Gene Symbol and Name	Dgkz, diacylglycerol kinase zeta
Gene Synonym(s)
Expressed Gene	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.
Strain of Origin	129X1/SvJ
Chromosome	2
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.
Mutations Made By	Gary Koretzky, University of Pennsylvania

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Immunology, Inflammation and Autoimmunity Research
- Immunodeficiency
  - T cell deficiency
- T Cell Receptor Signaling Defects

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Dgkz^tm1Gak/Dgkz^tm1Gak
involves: 129X1/SvJ * C57BL/6

cellular phenotype

increased T cell proliferation
- T cells are hyperresponsive to TCR stimulation

homeostasis/metabolism phenotype

abnormal interferon level
- than wild-type T cells
- a higher percentage of lymphocytic chriomeningitis virus-specific CD8+ T cells stimulated with major histocompatibility complex class I-specific peptide NP396 or the major histocompatibility complex class II-specific peptide GP61 express interferon-gamma than wild-type T cells

abnormal cytokine level
- bone marrow macrophages stimulated with LPS (a TLR4 ligand) or upon TLR3 activation with Poly(I:C) or TLR2 activation with Pam3CSK4 show reduced transcript levels of IL-6, TNF-alpha, and IL-1beta
- SCW-treated mice to induce arthritis show lower expression levels of proinflammatory cytokines IL-6, TNF-alpha, and IL-1beta
- CpG-administered mice to induce cytokine storm syndrome show lower IL-1beta, IL-6, and TNF-alpha mRNA levels in the liver

decreased circulating ferritin level
- mice administered the TLR9 ligand CpG to induce cytokine storm syndrome show lower serum ferritin levels

immune system phenotype

abnormal cytokine level
- bone marrow macrophages stimulated with LPS (a TLR4 ligand) or upon TLR3 activation with Poly(I:C) or TLR2 activation with Pam3CSK4 show reduced transcript levels of IL-6, TNF-alpha, and IL-1beta
- SCW-treated mice to induce arthritis show lower expression levels of proinflammatory cytokines IL-6, TNF-alpha, and IL-1beta
- CpG-administered mice to induce cytokine storm syndrome show lower IL-1beta, IL-6, and TNF-alpha mRNA levels in the liver

abnormal cytokine secretion
- bone marrow macrophages stimulated with LPS, Pam3CSK4, or CpG secrete lower levels of proinflammatory cytokines

decreased CD4-positive, alpha beta T cell number
- spleen and lymph nodes show a 5-10% decrease of Thy1+ cells in both CD4+ and CD8+ compartments, however CD4+ T cell levels are normal in the thymus

decreased CD8-positive, alpha-beta T cell number
- spleen and lymph nodes show a 5-10% decrease of Thy1+ cells in both CD4+ and CD8+ compartments, however CD8+ T cell numbers are normal in the thymus

decreased interleukin-6 secretion
- CpG-administered mice show decreased IL-6 serum levels compared to wild-type mice

decreased susceptibility to viral infection
- immune response to lymphocytic choriomeningitis virus infection is more robust than that observed in wild-type and mutants show a 50-70% reduction in viral titeres

increased B cell number
- spleen and lymph nodes show a compensatory increase of B220+ B cells to the decrease in T cells

increased T cell proliferation
- T cells are hyperresponsive to TCR stimulation

abnormal interferon level
- than wild-type T cells
- a higher percentage of lymphocytic chriomeningitis virus-specific CD8+ T cells stimulated with major histocompatibility complex class I-specific peptide NP396 or the major histocompatibility complex class II-specific peptide GP61 express interferon-gamma than wild-type T cells

abnormal macrophage physiology
- marker analysis indicates that bone marrow macrophages exhibit impaired M1 macrophage polarization

abnormal inflammatory response
- mice administered the TLR9 ligand CpG to induce cytokine storm syndrome show a similar increase in spleen and liver weight, decrease in WBC counts, areas of inflammatory infiltrates in the liver and percentage of F4/80+CD11b+ liver macrophages as in controls indicting that tissue infiltration by monocytes/macrophages is not affected, however serum ferritin levels are lower and production of inflammatory cytokines is reduced

abnormal T cell activation
- mice infected with lymphocytic chriomeningitis virus (LCMV) show a greater increase in total splenic CD8+ cells and a higher percentage of CD8+ cells with an activated phenotype 7 days after infection

abnormal T cell physiology
- TCR-induced phosphatidic acid production by T cells is inhibited compared with that of wild-type

decreased susceptibility to induced arthritis
- mice injected with streptococcal cell wall (SCW) fragments in the ankle to induce a local inflammatory response are protected from SCW-induced arthritis, showing less swelling and despite the presence of inflammatory infiltrate, protection from joint and cartilage disruption

decreased susceptibility to Riboviria infection
- immune response to lymphocytic choriomeningitis virus infection is more robust than that observed in wild-type and mutants show a 50-70% reduction in viral titeres

skeleton phenotype

decreased susceptibility to induced arthritis
- mice injected with streptococcal cell wall (SCW) fragments in the ankle to induce a local inflammatory response are protected from SCW-induced arthritis, showing less swelling and despite the presence of inflammatory infiltrate, protection from joint and cartilage disruption

hematopoietic system phenotype

increased T cell proliferation
- T cells are hyperresponsive to TCR stimulation

abnormal T cell physiology
- TCR-induced phosphatidic acid production by T cells is inhibited compared with that of wild-type

decreased CD4-positive, alpha beta T cell number
- spleen and lymph nodes show a 5-10% decrease of Thy1+ cells in both CD4+ and CD8+ compartments, however CD4+ T cell levels are normal in the thymus

abnormal T cell activation
- mice infected with lymphocytic chriomeningitis virus (LCMV) show a greater increase in total splenic CD8+ cells and a higher percentage of CD8+ cells with an activated phenotype 7 days after infection

abnormal macrophage physiology
- marker analysis indicates that bone marrow macrophages exhibit impaired M1 macrophage polarization

increased B cell number
- spleen and lymph nodes show a compensatory increase of B220+ B cells to the decrease in T cells

decreased CD8-positive, alpha-beta T cell number
- spleen and lymph nodes show a 5-10% decrease of Thy1+ cells in both CD4+ and CD8+ compartments, however CD8+ T cell numbers are normal in the thymus

The following phenotype relates to a compound genotype created using this strain

Genotype: Dgkz^tm1Gak/Dgkz^tm1Gak
B6.129X1-Dgkz

hematopoietic system phenotype

increased T cell number
- stimulation of CD8+ T cell-depleted splenocyte cultures in which Dgka production is inhibited results in 2-3 rounds of proliferation in mutant T cells; mutant T cells grow and divide like wild-type T cells that receive CD28 costimulation

immune system phenotype

abnormal lymphocyte anergy
- treatment with anti-CD3 and anti-CD28 which induces anergy in wild-type T cells does not affect mutant T cells which can still grow and produce IL-2

abnormal interleukin-2 secretion
- after stimulation with anti-CD3 and CTLA-4, wild-type T cells become anergic and produce very little IL-2, but mutant cells produce more IL-2; treatment of mutant cells with Dgka inhibitor results in levels of IL-2 production similar to those in anti-CD3-treated wild-type cells receiving CD28 costimulation
- treated wild-type cells receiving CD28 costimulation

increased T cell number
- stimulation of CD8+ T cell-depleted splenocyte cultures in which Dgka production is inhibited results in 2-3 rounds of proliferation in mutant T cells; mutant T cells grow and divide like wild-type T cells that receive CD28 costimulation

References

Zhong XP; Hainey EA; Olenchock BA; Jordan MS; Maltzman JS; Nichols KE; Shen H; Koretzky GA. 2003. Enhanced T cell responses due to diacylglycerol kinase zeta deficiency. Nat Immunol 4(9):882-90PubMed: 12883552 MGI: J:85149

Additional - Dgkz^tm1Gak related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Dgkz
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, homozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the B6.129X1-Dgkz^tm1Gak/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018861 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
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Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Dgkz<tm1Gak>

Related Products and Services

Frozen Mouse Embryo	B6.129X1-Dgkz<tm1Gak>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129X1-Dgkz<tm1Gak>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129X1-Dgkz<tm1Gak>/J Frozen Embryo	$3373.50
Frozen Mouse Embryo	B6.129X1-Dgkz<tm1Gak>/J Frozen Embryo	$3373.50

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The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

General Terms and Conditions

Questions about Terms of Use

Additional Use Restrictions Apply

Use of MICE by companies or for-profit entities requires a license prior to shipping.

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Dgkztm1Gak

Allele Symbol: Dgkztm1Gak

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Dgkztm1Gak/Dgkztm1Gakinvolves: 129X1/SvJ * C57BL/6

cellular phenotype

homeostasis/metabolism phenotype

immune system phenotype

skeleton phenotype

hematopoietic system phenotype

Genotype: Dgkztm1Gak/Dgkztm1GakB6.129X1-Dgkz

hematopoietic system phenotype

immune system phenotype

References

Additional - Dgkztm1Gak related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

Dgkz^tm1Gak

Allele Symbol: Dgkz^tm1Gak

Genotype: Dgkz^tm1Gak/Dgkz^tm1Gak
involves: 129X1/SvJ * C57BL/6

Genotype: Dgkz^tm1Gak/Dgkz^tm1Gak
B6.129X1-Dgkz

Additional - Dgkz^tm1Gak related