The dbl-PAC-Tg(SNCAA53T);Snca-/-; GbaL444P/+ mice harbor a Snca knockout allele, two transgenes encoding the human A53T-mutant alpha-synuclein associated with autosomal dominant Parkinson's disease and the L444P mutation in the Gba (glucosidase, beta, acid) gene associated with the alpha-synucleinopathy, Gaucher disease. Combining these two mouse models of Parkinson’s and Gaucher disease results in an exacerbation of the motor and gastrointestinal deficits found in the Parkinson’s A53T model alone.
Robert L Nussbaum, University of California San Francisco
Genetic Background | Generation |
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?+pN1
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Snca | synuclein, alpha |
Allele Type |
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Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Type |
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Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Hypomorph, Humanized sequence) | Gba | glucosidase, beta, acid |
The dbl-PAC-Tg(SNCAA53T);Snca-/-; GbaL444P/+ mice harbor a Snca knockout allele, two independently inserted transgenes encoding the human A53T-mutant alpha-synuclein associated with autosomal dominant Parkinson's disease and the L444P mutation in the Gba (glucosidase, beta, acid) gene associated with the alpha-synucleinopathy, Gaucher disease.
In mice homozygous for the two transgenes and the Snca KO, expression of endogenous mouse alpha-synuclein is abolished and replaced by alpha-synuclein*A53T from the four total insertions of the PAC- Tg(SNCAA53T) resulting in a Parkinson’s phenotype (see Jackson Laboratory Stock No. 010799). The addition of the heterozygous Gaucher mutation, L444P, reduces glucocerebrosidase (GBA) activity by 40%, reduces SNCA degradation, and results in the accumulation SNCA protein in culture. The enzyme GBA is involved in lysosomal protein degradation and thus in the lysosomal-autophagy pathway mainly responsible for SNCA degradation.
Heterozygous carriers of Gaucher disease have an increased risk for Parkinson’s disease.
14 to 15 month old mice exhibit an enteric nervous system dysfunction characterized by constipation and reduced whole gut transit time (males only). Motor activity is impaired as well as exploratory behavior, and endurance and coordination on the rotarod. Accumulation of human pSer129 SNCA is observed earlier (15 months) than in the A53T mice, although there is no increase in absolute SNCA levels.
Combining these two mouse models of Parkinson’s and Gaucher disease results in an exacerbation of the motor and gastrointestinal deficits found in the Parkinson’s A53T model alone.
The PAC-Tg(SNCAA53T) transgene was designed using the 146 kb RPCI-1 human male P1 artificial chromosome (PAC) clone 27M07, containing the entire human SNCA (synuclein, alpha (non A4 component of amyloid precursor)) gene modified to have the A53T human mutation. This transgene was microinjected into the pronuclei of fertilized FVB/N ova. Each transgenic founder mouse was bred to FVB/N wildtype mice to generate the individual founder lines (1 and 2).
The targeting vector for the Snca knockout allele was designed to replace exons 4 and 5 with a reverse-oriented neomycin resistance cassette. The construct was electroporated into 129S6/SvEvTac-derived TC-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and the resulting chimeric mice were crossed to 129S6/SvEvTac.
The transgenic dbl-PAC-Tg(SNCAA53T) strains and the Snca-/- strain were combined to form a homozygous double transgenic, homozygous Scna KO strain maintained on a mixed FVB/N;129S6/SvEvTac background. The triple mutant is distributed by The Jackson Laboratory as Stock No. 010799).
The targeting vector for the Gba L444P knockin allele was designed to insert a proline to leucine substitution mutation at amino acid position 444 in exon 10. The construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and the resulting chimeric mice were crossed to C57BL/6. The GbaL444P mutant is distributed by MMRRC as Stock No. 000117. The triple SCNA mutant was crossed to GbaL444P mice and a colony was generated by random intrastrain matings that is homozygous for both transgenes and the Scna KO and heterozygous for the GbaL444P allele.
Expressed Gene | SNCA, synuclein alpha, human |
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Site of Expression | |
Expressed Gene | SNCA, synuclein alpha, human |
Site of Expression | |
Site of Expression |
Allele Name | targeted mutation 1, Robert L Nussbaum |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Scna-; Snca- |
Gene Symbol and Name | Snca, synuclein, alpha |
Gene Synonym(s) | |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | Exons 4 and 5 were replaced with a neomycin selection cassette inserted by homologous recombination. While the separation of Western blot results by 2D-PAGE showed various isoforms in total brain extract obtained from wild-type mice, protein was undetected in samples obtained from homozygous mutant mice. |
Allele Name | transgene insertion 1, Robert L Nussbaum |
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Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | hSNCAA53T; PAC-Tg(SCNAA53T) |
Gene Symbol and Name | Tg(SNCA*A53T)1Nbm, transgene insertion 1, Robert L Nussbaum |
Gene Synonym(s) | |
Promoter | SNCA, synuclein alpha, human |
Expressed Gene | SNCA, synuclein alpha, human |
Strain of Origin | FVB/N |
Chromosome | UN |
Molecular Note | To generate the PAC-Tg(SNCAA53T) transgene, the 146 kb RPCI-1 human male P1 artificial chromosome (PAC) clone 27M07, containing the entire human SNCA (synuclein, alpha (non A4 component of amyloid precursor)) gene and 34 kb of its upstream region, was modified to have the A53T human mutation associated with autosomal dominant Parkinson's disease. This transgene was microinjected into the pronuclei of fertilized FVB/N ova. One transgenic insertion is present in the genome. |
Allele Name | transgene insertion 2, Robert L Nussbaum |
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Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | hSCNAA53T; PAC-Tg(SCNAA53T) |
Gene Symbol and Name | Tg(SNCA*A53T)2Nbm, transgene insertion 2, Robert L Nussbaum |
Gene Synonym(s) | |
Promoter | SNCA, synuclein alpha, human |
Expressed Gene | SNCA, synuclein alpha, human |
Strain of Origin | FVB/N |
Chromosome | UN |
Molecular Note | To generate the PAC-Tg(SNCAA53T) transgene, the 146 kb RPCI-1 human male P1 artificial chromosome (PAC) clone 27M07, containing the entire human SNCA (synuclein, alpha (non A4 component of amyloid precursor)) gene and 34 kb of its upstream region, was modified to have the A53T human mutation associated with autosomal dominant Parkinson's disease. This transgene was microinjected into the pronuclei of fertilized FVB/N ova. One transgenic insertion is present in the genome. |
Allele Name | targeted mutation 1, Richard L Proia |
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Allele Type | Targeted (Hypomorph, Humanized sequence) |
Allele Synonym(s) | GbaL444P; L444P |
Gene Symbol and Name | Gba, glucosidase, beta, acid |
Gene Synonym(s) | |
Strain of Origin | 129S4/SvJae |
Chromosome | 3 |
Molecular Note | A proline to leucine substitution mutation at amino acid position 444 is associated with Gaucher disease type 3 in humans. This mutation was introduced in mice using a single insertion mutagenesis procedure. The level of gene expression in brain of mutant mice is similar to wild-type as determined by Northern blot analysis, but enzyme assays with liver, brain, and skin extracts demonstrate that enzyme activity is 20% that of wild-type. |
The colony is maintained as homozygous for both transgenes and the Snca KO and heterozygous for the GbaL444P allele. Mice homozygous for GbaL444P die shortly after birth.
When using the GbaL444P; Snca-; dbl-PAC-Tg(SNCAA53T) mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37633 in your Materials and Methods section.
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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