These mice express the mutant D409V GBA protein and possess loxP sites flanking exons 6 through 8 of the Gba (glucosidase, beta, acid) gene. They have applications in studies related to Parkinson's disease, Gaucher disease and synucleinopathies.
Kuldip Dave, The Michael J. Fox Foundation
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 the mutant D427V Gba protein, which corresponds to the D409V mutation in the mature human GBA protein, and possess loxP sites flanking exons 6 through 8. Mice that are heterozygous for the targeted mutation are viable and fertile. Homozygous Gba D409V knock-in mice show a significant reduction in Gba protein level and glucocerebrosidase activity, and a significant increase in glucosylsphingosine in both brain and liver tissues.
Heterozygotes demonstrate significant cognitive impairment by the age of 12 months, as determined by both Morris water maze and Y-maze behavioral tests (PMID: 31299418).
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.
Preliminary characterization was presented in a poster at the 2015 Society for Neuroscience conference.
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 (see SNP results below). These mice no longer carry the FLP recombinase allele. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6N (Stock No. 005304) at least once to establish the colony.
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 the 27 markers throughout the genome suggested a C57BL/6 genetic background, 4 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.
|Allele Name||targeted mutation 1.1, The Michael J Fox Foundation|
|Allele Type||Targeted (Humanized sequence, Not Specified)|
|Allele Synonym(s)||Gbatm1(D427V)Mjff; Gba1D409V|
|Gene Symbol and Name||Gba, glucosidase, beta, acid|
|Promoter||Gba, glucosidase, beta, acid, mouse, laboratory|
|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.|
When maintaining a live colony, heterozygous mice may be bred together, to wild type mice from the colony or to C57BL/6NJ inbred mice (Stock No. 005304).
When using the Gba D409V KI mouse strain in a publication, please cite the originating article(s) and include JAX stock #019106 in your Materials and Methods section.