TgDNPKR mice express a catalytically-null/dominant-negative (K296R) mutant variant of human PKR (DNPKR) exclusively in hematopoietic cells/tissues under control of the mouse HS21/45-vav regulatory elements. This DNPKR expression results in loss of PKR activity in bone marrow cells and increased hematopoietic stem/progenitor cell populations, as well as bone marrow cell-resistance to apoptosis-inducing cell stress. These TgDNPKR mice may be useful in studying how loss of PKR activity may contribute to increased growth and malignancy.
W. Stratford May, University of Florida
Genetic Background | Generation |
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Allele Type |
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Transgenic (Inserted expressed sequence, Humanized sequence) |
Protein kinase R (PKR) is an interferon (IFN)-inducible, double-stranded RNA-activated kinase that initiates apoptosis in response to cellular stress and inhibits DNA damage response signaling. Loss of the PKR expression/activity has been associated with increased growth of human breast carcinoma, non-small cell lung cancer, B-cell chronic lymphocytic leukemia, and T-cell acute lymphoblastic leukemia, suggesting that the loss of PKR activity may contribute to increased growth and malignancy.
These vav-DNPKR transgenic mice (TgDNPKR) have the mouse HS21/45-vav regulatory elements directing expression of a catalytically-null/dominant-negative (K296R) mutant variant of human PKR to hematopoietic cells/tissues. High DNPKR expression is reported in thymus, spleen and bone marrow (BM) compared with non-hematopoietic tissues (small intestine, liver or kidney) as demonstrated by real-time quantitative PCR and western blotting. This DNPKR expression results in loss of PKR activity in BM cells and increased hematopoietic stem/progenitor cell populations (HSPCs), as well as BM cell-resistance to apoptosis-inducing cell stress.
Specifically, TgDNPKR mice have no significant BM abnormalities reported to date (May 2015). Compared with non-transgenic mice, the TgDNPKR mice show increased numbers of HSPCs in association with increased peripheral blood cell counts (WBC, platelet and hemoglobin levels) as the mice age. Also, TgDNPKR mice show elevated numbers of mature myeloid cell populations within the BM, more robust hematopoietic colony-forming capacity, and improved BM cell survival to various stresses that mediate cell death/apoptosis (including hematopoietic growth factor deprivation, treatment with inflammatory cytokines or irradiation).
Hemizygous TgDNPKR mice are viable and fertile. To date (July 2015), it has not been attempted to make this strain homozygous.
The phenotype of hemizygous TgDNPKR mice is in contrast to hemizygous mice expressing human wildtype (catalytically-active) PKR from the same HS21/45-vav regulatory elements (TgPKR; Stock No. 027460). Those TgPKR mice have overexpression of PKR resulting in significantly defective hematopoiesis, BM failure and increased sensitivity to apoptosis-inducing cell stress.
The vav-DNPKR transgene was designed by Dr. W. Stratford May Jr (University of Florida) with the mouse vav 1 oncogene promoter/control regions (Vav1; from the HS21/45-vav vector) directing expression of a catalytically-null/dominant-negative (K296R) mutant variant of human eukaryotic translation initiation factor 2-alpha kinase 2 cDNA sequence (EIF2AK2K296R or DNPKR).
Specifically, the transgene has (from 5' to 3') the two proximal upstream Vav1 DNase-I hypersensitive sites (HS21), an SV40 intron, the human DNPKR cDNA sequence (EIF2AK2 nucleotides 886-888 changed from AAA [lysine] to CGT [arginine]), an SV40 polyA signal and Vav1 intron 1 (containing both proximal HS sites (HS45)).
This 8.5 kbp vav-DNPKR transgene was microinjected into the pronuclei of C57BL/6J eggs, and then founder animals were bred to C57BL/6J mice for germline transmission. A single transgenic (TgDNPKR) founder lines was generated. TgDNPKR mice were then bred with C57BL/6J wildtype mice for more than seven generations prior to sending to The Jackson Laboratory Repository in 2015.
Upon arrival, males were used to cryopreserve sperm. To establish the living TgDNPKR mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).
Expressed Gene | EIF2AK2, eukaryotic translation initiation factor 2 alpha kinase 2, human |
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Site of Expression |
Allele Name | transgene insertion 1, W May |
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Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | TgDNPKR |
Gene Symbol and Name | Tg(Vav1-EIF2AK2*K296R)1Wsmay, transgene insertion 1, W May |
Gene Synonym(s) | |
Promoter | Vav1, vav 1 oncogene, mouse, laboratory |
Expressed Gene | EIF2AK2, eukaryotic translation initiation factor 2 alpha kinase 2, human |
Strain of Origin | C57BL/6 |
Chromosome | UN |
Molecular Note | The transgene consists of mouse vav 1 oncogene promoter/control regions directing expression of a catalytically-null/dominant-negative (K296R) mutant variant of human eukaryotic translation initiation factor 2-alpha kinase 2 cDNA sequence. |
When maintaining a live colony, hemizygous mice may be bred to wildtype (noncarrier) siblings or to C57BL/6J inbred mice (Stock No. 000664). To date (July 2015), it has not been attempted to make this strain homozygous.
When using the TgDNPKR mouse strain in a publication, please cite the originating article(s) and include JAX stock #027461 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous or non carrier for Tg(Vav1-EIF2AK2*K296R)1Wsmay |
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