This triple knockout strain is useful in studies of dopaminergic neuron physiology, regulation of striatal dopamine and serotonin, and Parkinson's disease.
Matthew Goldberg, University of Texas Southwestern Medical Center at Dallas
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Prkn | parkin RBR E3 ubiquitin protein ligase |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Park7 | Parkinson disease (autosomal recessive, early onset) 7 |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Null/Knockout) | Gpx1 | glutathione peroxidase 1 |
In the brains of patients with Parkinson's disease, the antioxidant enzyme glutathione peroxidase 1 activity level is diminished. Mutations in human PARK7 and PARK2 resulting loss of function have been linked to types of autosomal recessive Parkinson's disease. These mice carry targeted mutations for the Park7, Gpx1 and Park2 genes.
Mice that are homozygous for all 3 of the targeted mutations are viable and fertile.
In triple mutant mice, 12 to 18 months of age, striatal dopamine and serotonin levels are elevated, but with normal nigral neuron numbers. Parkin-/- DJ-1-/- Gpx1-/- exhibit enhanced motor behavior and are able to outperform wildtype controls in the rotarod test.
During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
For the Park7 A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While 26 of the 27 markers throughout the genome suggested a C57BL/6 genetic background, 2 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
& Women's Hospital, Harvard University) containing a PGK-neomycin cassette was used to disrupt exon 2. The construct was electroporated into (C57BL/6 x 129)F1 derived MKV6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission.
The mice were backcrossed to C57BL/6J for 10 generations.
For the Gpx1tm1Ysh allele, a targeting vector designed by Dr. Ye-Shih Ho (Wayne State University) containing a NEO cassette was used to disrupt exon 2. The construct was electroporated into 129 derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were tested for germline transmission.
The mice were crossed for approximately 6 generations to C57BL/6. Dr. Matthew Goldberg obtained the mice and backcrossed them to C57BL/6J for 3 or 4 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background.
For the Park2
To generate this triple mutant strain, the Park7 and Park2 knockout lines were bred and intercrossed to obtain double knockout mice. The double knockout mice were then crossed to the Gpx1 knockout mice for 2 generations, and then intercrossed to obtain mice homozygous for all 3 alleles, Parkin-/- DJ-1-/- Gpx1-/- (See SNP note below). Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Allele Name | targeted mutation 1, Jie Shen |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Park2tm1Shn; parkin - |
Gene Symbol and Name | Prkn, parkin RBR E3 ubiquitin protein ligase |
Gene Synonym(s) | |
Strain of Origin | 129S4/SvJae |
Chromosome | 17 |
Molecular Note | Exon 3 was replaced in-frame by the coding sequence for EGFP followed by a PGK-neomycin cassette. RT-PCR analysis indicated that exon 2 spliced to exon 4 in transcripts thus skipping exon 3 entirely. This results in a frame shift and a premature stop codon in exon 5. Western blot analysis using antibody specific to C-terminal sequences indicated the absence of gene product. |
Mutations Made By | Jie Shen, Harvard Med Sch/Brigham Women's Hosp |
Allele Name | targeted mutation 1, Jie Shen |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | DJ-1- |
Gene Symbol and Name | Park7, Parkinson disease (autosomal recessive, early onset) 7 |
Gene Synonym(s) | |
Strain of Origin | (C57BL/6 x 129)F1 |
Chromosome | 4 |
Molecular Note | Exon 2 was replaced with a pgk-neo cassette. Western blot failed to detect protein in mutant mice. |
Mutations Made By | Jie Shen, Harvard Med Sch/Brigham Women's Hosp |
Allele Name | targeted mutation 1, Ye-Shih Ho |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Gpx1-; GPX1 KO; GpX-1 KO; GSHPx-1-deficient |
Gene Symbol and Name | Gpx1, glutathione peroxidase 1 |
Gene Synonym(s) | |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 9 |
Molecular Note | Insertion of a neomycin resistance cassette disrupted exon 2 of the gene. Northern blots of total RNA isolated from brain, heart, kidney, liver, and lung of homozygous mutant mice showed no detectable transcript of the targeted gene when probed with an a gene-specific cDNA probe. |
When maintaining a live colony, these mice can be bred as homozygotes for each targeted mutation.
When using the Parkin -/-DJ-1-/-Gpx1-/- mouse strain in a publication, please cite the originating article(s) and include JAX stock #023968 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Homozygous for Park7<tm1Shn>, Homozygous for Gpx1<tm1Ysh>, Homozygous for Park2<tm1Shn> |
Frozen Mouse Embryo | B6.Cg-Park7<tm1Shn> Gpx1<tm1Ysh> Prkn<tm1Shn> Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Park7<tm1Shn> Gpx1<tm1Ysh> Prkn<tm1Shn> Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Park7<tm1Shn> Gpx1<tm1Ysh> Prkn<tm1Shn> Frozen Embryos | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Park7<tm1Shn> Gpx1<tm1Ysh> Prkn<tm1Shn> Frozen Embryos | $3373.50 |
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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.
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