Transgenic hN370S mice express a human GBA (glucosidase, beta, acid) gene modified to include the N370S (N409S) substitution associated with Gaucher disease on a Gba-null background. This strain may be useful in studies of the mutations underlying Gaucher disease.
Lorne A Clarke, University of British Columbia
The hN370S transgene uses a CAG promoter to direct expression of the human GBA (glucosidase, beta, acid) gene modified to incorporate an asparagine to serine substitution at residue 409 (also described as N370S). Mutations in GBA, particularly N370S and L444P, are associated with Gaucher disease, a storage disorder caused by the absence of acid B-glucosidase, a lysosomal hydrolase. When mice carrying the hN370S transgene are combined with mice homozygous for the Gba null allele (Gbatm1.1Clk), the transgene prevents neonatal lethality in progeny. In the first 24 hours, pups exhibit wrinkled and dried skin, but by 48 hours, pups resemble normal littermates. In addition, hN370S-tg4 Gba-null mice exhibit increases in glucosylceramide and glucosylsphingosine levels in the spleen and liver. Glucosylsphingosine levels also are increased in the central nervous system. This strain may be useful in studies of the mutations underlying Gaucher disease.
This strain is one of two Gba null strains carrying modified GBA transgenes - B6;129-Gbatm1.1Clk Tg(CAG-GBA*L483P)#Clk/Mmjax - Stock No. 050598.
The targeting construct for mouse Gba (glucosidase, beta, acid) contains loxP sites inserted upstream of exon 9 and downstream of exon 11 followed by a FRT-flanked neomycin cassette. Transient expression of FLP recombinase removed the neomycin cassette. The construct was electroporated into (129X1/SvJ x 129S1/Sv)-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred. Mice were crossed to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J to remove exons 9-11 and generate the null allele.
The transgenic construct contains the human GBA (glucosidase, beta, acid) gene under the direction of CMV enhancer/chicken beta-actin promoter, rabbit beta-globin intron sequence and the rabbit globin polyA site (pCAGEN) promoter. GBA was modified by site-directed mutagenesis (c.1226A to G) to harbor an asparagine to serine mutation at residue 409 (also described as N370S). The linearized construct was microinjected into fertilized C57BL/6xCBA oocytes. Founders were crossed to mice on a mixed C57BL/6 and 129Sv background. The founder mouse was subsequently established and bred to mice carrying Gbatm1.1Clk on a mixed background (C57BL/6, 129 and FVB/N) . Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.
|Allele Name||targeted mutation 1.1, Lorne A Clarke|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||Gbatm1.1Clk; targeted mutation 1.1, Lorne A Clarke|
|Gene Symbol and Name||Gba, glucosidase, beta, acid|
|Gene Synonym(s)||betaGC; GC; GCase; GCB; GBA1; GLUC; glucocerebrosidase|
|Strain of Origin||(129X1/SvJ x 129S1/Sv)F1-Kitl+|
|Molecular Note||Cre mediated recombination removed floxed sequence containing exons 9-11, leaving metataxin exons 8-4.|
|Allele Name||transgene insertion, Lorne A Clarke|
|Allele Type||Transgenic (Inserted expressed sequence, Humanized sequence)|
|Allele Synonym(s)||Tg(CAG-GBA*N409S)#Clk; transgene insertion, Lorne A Clarke|
|Gene Symbol and Name||Tg(CAG-GBA*N409S)#Clk, transgene insertion, Lorne A Clarke|
|Strain of Origin||C57BL/6 x CBA|
|Molecular Note||The transgenic construct contains the human GBA gene under the direction of CMV enhancer/chicken beta-actin promoter, rabbit beta-globin intron sequence and the rabbit globin polyA site (pCAGEN) promoter. GBA was modified by site-directed mutagenesis (c.1226A >G) to harbor an asparagine to serine mutation at residue 409 (referred to as N370S).|
When maintaining a live colony, Gbatm1.1Clk homozygous and Tg hemizygous mice may be bred to mice homozygous for Gbatm1.1Clk . The donating investigator has not attempted to generate mice homozygous for the transgene.
When using the Gba-/- hN370S mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #50597 in your Materials and Methods section.
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