Overview

Also Known As:Gba^L444P; Snca^-; dbl-PAC-Tg(SNCA^A53T)

The dbl-PAC-Tg(SNCA^A53T);Snca^-/-; Gba^L444P/+ 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.

Donating Investigator

Robert L Nussbaum, University of California San Francisco

Genetic overview

Genetic Background	Generation
	?+pN1 (2020-04-20 00:00:00)

Snca^tm1Nbm

Allele Type	Gene Symbol	Gene Name
Targeted (Null/Knockout)	Snca	synuclein, alpha

Tg(SNCA*A53T)1Nbm

Allele Type
Transgenic (Inserted expressed sequence, Humanized sequence)

Tg(SNCA*A53T)2Nbm

Allele Type
Transgenic (Inserted expressed sequence, Humanized sequence)

Gba^tm1Rlp

Allele Type	Gene Symbol	Gene Name
Targeted (Hypomorph, Humanized sequence)	Gba	glucosidase, beta, acid

View Genetics

Research Applications

Internal/Organ Research
Neurobiology Research

View All Research Applications

Details

Detailed Description

The dbl-PAC-Tg(SNCA^A53T);Snca^-/-; Gba^L444P/+ 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(SNCA^A53T) 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.

Development

The PAC-Tg(SNCA^A53T) 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(SNCA^A53T) 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 Gba^L444P mutant is distributed by MMRRC as Stock No. 000117. The triple SCNA mutant was crossed to Gba^L444P mice and a colony was generated by random intrastrain matings that is homozygous for both transgenes and the Scna KO and heterozygous for the Gba^L444P allele.

Expression Data

Expressed Gene	SNCA, synuclein alpha, human
Site of Expression
Expressed Gene	SNCA, synuclein alpha, human
Site of Expression
Site of Expression

Control Suggestions

A control for this strain is available; see STOCK Gba^tm1Rlp Snca^tm1Nbm Tg(SNCA)1Nbm/Mmjax MMRRC Stock Number 037801.

Additional Information

Considerations for Choosing Controls

Selected References

Fishbein I; Kuo YM; Giasson BI; Nussbaum RL. 2014. Augmentation of phenotype in a transgenic Parkinson mouse heterozygous for a Gaucher mutation. Brain 137(Pt 12):3235-47PubMed: 25351739 MGI: J:217865

View All References

Genetics

Snca^tm1Nbm

Allele Symbol: Snca^tm1Nbm

Allele Name	targeted mutation 1, Robert L Nussbaum
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.

Tg(SNCA*A53T)1Nbm

Allele Symbol: Tg(SNCA*A53T)1Nbm

Allele Name	transgene insertion 1, Robert L Nussbaum
Allele Type	Transgenic (Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	hSNCA^A53T; PAC-Tg(SCNA^A53T)
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.

Tg(SNCA*A53T)2Nbm

Allele Symbol: Tg(SNCA*A53T)2Nbm

Allele Name	transgene insertion 2, Robert L Nussbaum
Allele Type	Transgenic (Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	hSCNA^A53T; PAC-Tg(SCNA^A53T)
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.

Gba^tm1Rlp

Allele Symbol: Gba^tm1Rlp

Allele Name	targeted mutation 1, Richard L Proia
Allele Type	Targeted (Hypomorph, Humanized sequence)
Allele Synonym(s)	Gba^L444P; 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.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Internal/Organ Research
- Gastrointestinal Defects
Neurobiology Research
- Parkinson's Disease
  - synuclein mutants

Mammalian Phenotype Terms by Genotype

References

Fishbein I; Kuo YM; Giasson BI; Nussbaum RL. 2014. Augmentation of phenotype in a transgenic Parkinson mouse heterozygous for a Gaucher mutation. Brain 137(Pt 12):3235-47PubMed: 25351739 MGI: J:217865

Additional - Gba^tm1Rlp related

Additional - Snca^tm1Nbm related

Additional - Tg(SNCA*A53T)1Nbm related

Additional - Tg(SNCA*A53T)2Nbm related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- QPCR:Tg(SNCAcDNA)-qPCR
- Standard PCR:Tg(SNCA)129Mjff
- Sanger sequencing:Gba
- Separated PCR:Snca
- Standard PCR:Tg(SNCA)
- Genotyping resources and troubleshooting
Breeding Considerations

The colony is maintained as homozygous for both transgenes and the Snca KO and heterozygous for the Gba^L444P allele. Mice homozygous for Gba^L444P die shortly after birth.
- Additional Breeding and Husbandry Support
Citation
When using the Gba^L444P; Snca^-; dbl-PAC-Tg(SNCA^A53T) mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #37633 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

FGB29 (Standard)

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

See MMRRC for Additional Conditions of Distribution

Questions about Terms of Use

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Also Known As:GbaL444P; Snca-; dbl-PAC-Tg(SNCAA53T)

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Sncatm1Nbm

Allele Symbol: Sncatm1Nbm

Tg(SNCA*A53T)1Nbm

Allele Symbol: Tg(SNCA*A53T)1Nbm

Tg(SNCA*A53T)2Nbm

Allele Symbol: Tg(SNCA*A53T)2Nbm

Gbatm1Rlp

Allele Symbol: Gbatm1Rlp

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Gbatm1Rlp related

Additional - Sncatm1Nbm related

Additional - Tg(SNCA*A53T)1Nbm related

Additional - Tg(SNCA*A53T)2Nbm related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

Also Known As:Gba^L444P; Snca^-; dbl-PAC-Tg(SNCA^A53T)

Snca^tm1Nbm

Allele Symbol: Snca^tm1Nbm

Gba^tm1Rlp

Allele Symbol: Gba^tm1Rlp

Additional - Gba^tm1Rlp related

Additional - Snca^tm1Nbm related