Transgenic hL444P mice express a human GBA (glucosidase, beta, acid) gene modified to include the L444P (L483P) 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 hL444 transgene uses a CAG promoter to direct expression of the human GBA (glucosidase, beta, acid) gene modified to incorporate an leucine to proline substitution at residue 483 (described as L444P). 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 hL444P transgene are combined with mice homozygous for the Gba null allele (Gbatm1.1Clk), the transgene prevents neonatal lethality in progeny. By 52 weeks of age, hL444P/Gba-null mice develop pearly nodular splenic lesions. Gaucher cells are found in the spleen beginning at 45 weeks and increase progressively in number with age. Gaucher cells are not found in the liver, lung or bone marrow. hL444P 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*N409S)#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 an 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 the CMV enhancer/chicken beta-actin promoter, rabbit B-globin intron sequence and the rabbit globin polyA site (pCAGEN) promoter. GBA was modified by site-directed mutagenesis (c.1448T to C) to harbor an leucine to proline mutation at residue 483 (described as L444P). The linearized construct was microinjected into fertilized oocytes on an unspecified background. Founders were crossed to mice on a mixed C57BL/6 and 129Sv background. hL444P mice were bred to 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 8, Lorne A Clarke|
|Allele Type||Transgenic (Inserted expressed sequence, Humanized sequence)|
|Allele Synonym(s)||transgene insertion 8, Lorne A Clarke; Tg(CAG-GBA*L483P)8Clk|
|Gene Symbol and Name||Tg(CAG-GBA*L483P)8Clk, transgene insertion 8, Lorne A Clarke|
|Strain of Origin||Not Specified|
|Molecular Note||The transgenic construct contains the human GBA gene under the direction of the CMV enhancer/chicken beta-actin promoter, rabbit B-globin intron sequence and the rabbit globin polyA site (pCAGEN) promoter. GBA was modified by site-directed mutagenesis (c.1448T>C) to harbor an leucine to proline mutation at residue 483 (described as L444P).|
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-/- hL444P mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #50598 in your Materials and Methods section.
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