C57BL/6N-Gba^tm1.1Mjff Tg(Thy1-SNCA)15Mjff/J

Stock number: 029124
Product name: GBA1 D409V KI x mThy1-hSNCA
Research areas: Neurobiology Research

Description

These mice carry the Gba^tm1.1Mjff allele and the Tg(Thy1-SNCA)15Mjff transgene. They have applications related to Parkinson's disease, Gaucher disease and synucleinopathies, and neurodegeneration elicited by the aggregation of SNCA.

Detailed description

Mutations in human GBA (glucosidase, beta, acid) can cause Gaucher disease, an autosomal recessive lysosomal (lipid) storage disease, and have been associated with late onset Parkinson's disease and Lewy body dementia.

These mice express a transgene containing human SNCA under the direction of a mouse Thy-1 promoter, and the mutant D427V GBA protein, which corresponds to the D409V mutation in the mature GBA protein. In addition, these mice possess loxP sites flanking exons 6 through 8 of the Gba gene. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exons 6 through 8 deleted in the cre-expressing tissues and will no longer express the mutant D427V protein.

Mice that are heterozygous for the Gba^tm1.1Mjff allele and hemizygous for the Tg(Thy1-SNCA)15Mjff transgene are viable and fertile. Homozygous fertility for the Gba^tm1.1Mjff allele or transgene has not been tested (June, 2016). As the mice are characterized, we will modify the strain description and add phenotype data.

Development

Mice carrying the Gba^tm1.1Mjff allele were bred to mice carrying the Tg(Thy1-SNCA)15Mjff transgene to generate this double mutant line.

For the Gba^tm1.1Mjff allele, a targeting vector containing a FRT site flanked NEO cassette, and a F3 FRT site flanked PurO cassette was used to introduce the D427V point mutation into exon 10. The FRT site flanked NEO cassette was inserted into intron 5 and the F3 FRT site flanked PurO cassette was inserted into intron 8. In addition, loxP sites were inserted to flank exons 6 through 8. The construct was electroporated into C57BL/6NTac derived Art B6 3.6 embryonic stem (ES) cells. Correctly targeted ES cells were injected into BALB/c blastocysts. The resulting chimeric animals were crossed to FLP recombinase expressing females on the C57BL/6NTac background, C57BL/6NTac-Tg(CAG-Flpe)2Art, to remove the selection cassettes. These mice no longer carry the FLP recombinase allele. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6NJ (Stock No. 005304) at least once to establish the colony.

The mThy1-hSNCA transgene consists of the human a-synuclein (SNCA) gene driven by a mouse thymus cell antigen 1 (Thy-1) promoter. The construct was microinjected into fertilized C57BL/6NTac oocytes and males from founder line 15 were bred to C57BL/6NTac females to establish the colony. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ inbred mice (Stock No. 005304) for at least one generation.

Allele

Symbol: Gba^tm1.1Mjff
Name: targeted mutation 1.1, The Michael J Fox Foundation
MGI accession ID: MGI:5461046
Generation method: Targeted
Attributes: Humanized sequence; Not Specified
Marker symbol: Gba
Marker name: glucosidase, beta, acid
Chromosome: 3
Strain of origin: C57BL/6NTac
Molecular note: A targeting vector containing a FRT site flanked NEO cassette, and a F3 FRT site flanked PurO cassette was used to introduce the D427V point mutation into exon 10. D427V corresponds to the human D409V mutation. The FRT site flanked NEO cassette was inserted into intron 5 and the F3 FRT site flanked PurO cassette was inserted into intron 8. In addition, loxP sites were inserted to flank exons 6 through 8. Flp-mediated recombination removed the neo cassette.

Allele

Symbol: Tg(Thy1-SNCA)15Mjff
Name: transgene insertion 15, The Michael J Fox Foundation
MGI accession ID: MGI:5308782
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(Thy1-SNCA)15Mjff
Marker name: transgene insertion 15, The Michael J Fox Foundation
Chromosome: 11
Strain of origin: C57BL/6NTac
Expressed genes: SNCA,synuclein alpha,human
Molecular note: The transgene consists of the human alpha-synuclein (SNCA) gene driven by a mouse thymus cell antigen 1 (Thy-1) promoter. Line 15 was generated. Transgene insertion occurred on Chr 11, causing a 38 Kb deletion.