When Myh9flox mice are crossed with Pod::Cre mice resulting offspring will have exon 1 deleted in kidney podocytes. Double mutant mice on the C57BL/6 background do not develop spontaneous proteinuria or renal insufficiency, however, when subjected to environmental stress via doxyrubicin hydrochloride (Adriamycin), Myh9 podocyte-deleted mice develop albuminuria, focal and segmental glomerulosclerosis, foot process fusion and foot process effacement.
Duncan Johnstone, University of Pennsylvania
Genetic Background | Generation |
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Allele Type |
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Transgenic (Recombinase-expressing) |
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed), No functional change) | Myh9 | myosin, heavy polypeptide 9, non-muscle |
Myh9 (myosin, heavy chain 9, non-muscle) encodes a nonmuscle myosin IIA heavy chain involved in several functions including cytokinesis, cell motility and maintenance of cell shape. Genome wide association studies associate single nucleotide polymorphisms (SNPs) in Myh9 with chronic kidney disease in African-Americans. Myh9flox mutant mice possess loxP sites flanking exon 1. Homozygous mice are viable and fertile.
The podocin nephrosis 2 homolog (NPHS2) promoter directs expression of Cre recombinase to podocytes in the newborn and adult kidneys. Expression is minimal or undetectable in all other tissues tested and in 8.5 dpc embryos. Mice homozygous for Pod::Cre are viable and fertile.
When Myh9flox mice are crossed with Pod::Cre mice resulting offspring will have exon 1 deleted in kidney podocytes. Double mutant mice on the C57BL/6 background do not develop spontaneous proteinuria or renal insufficiency, however, when subjected to environmental stress via doxyrubicin hydrochloride (Adriamycin), Myh9 podocyte-deleted mice develop albuminuria, focal and segmental glomerulosclerosis, foot process fusion and foot process effacement.
This strain is unpublished on the FVB background. The donating investigator indicates that FVB mice are more sensitive to glomerular disease than C57BL/6 mice.
The Myh9tm1.1Gac targeting vector was designed to insert a loxP site upstream of exon 1 followed by a loxP-flanked neomycin resistance (neo) cassette downstream of exon 1 in the Myh9 gene. The construct was electroporated into 129/Sv-derived embryonic stem (ES) cells. Transient Cre expression in targeted cells excised the neo cassette leaving exon 1 flanked by loxP sites. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice for at least 5 generations.
The Pod::Cre (or 2.5P-Cre) transgene was designed with 2.5 kb fragment of the human podocin nephrosis 2 homolog (NPHS2) promoter/enhancer region followed by a Cre recombinase cassette. The transgene was microinjected into the pronuclei of fertilized oocytes from (C57BL/6 x SJL)F2 mice. Founder line 295 was established and used to generate mutant mice. The resulting transgenic mice were subsequently bred to wild-type C57BL/6 mice for an unknown number of generations.
Both strains were transferred to the University of Pennsylvania and intercrossed. Double mutant mice (Pod::Cre;Myh9f) were backcrossed to FVB/NJ mice by marker assisted selection (speed congenic) for 4 generations and then by traditional backcross for 10 generations.
The donating investigator maintains the strain by Pod::Cre/+ Myh9f/+ x FVB/NJ or Myh9f/f. Upon arrival, mice were bred to FVB/NJ for at least 1 generation to establish the colony.
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
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Site of Expression | cre recombinase expression in podocytes within kidney glomeruli. |
Site of Expression |
Allele Name | transgene insertion 295, Lawrence B Holzman |
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Allele Type | Transgenic (Recombinase-expressing) |
Allele Synonym(s) | 2.5P-Cre; NPHS2.Cre; pod-Cre; podocin-Cre; PodoCre; Tg(NPHS2-cre)1Lbh |
Gene Symbol and Name | Tg(NPHS2-cre)295Lbh, transgene insertion 295, Lawrence B Holzman |
Gene Synonym(s) | |
Promoter | NPHS2, NPHS2, podocin, human |
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
Site of Expression | cre recombinase expression in podocytes within kidney glomeruli. |
Strain of Origin | (C57BL/6 x SJL)F2 |
Chromosome | UN |
General Note | 24 transgenic founders were created. Phenotypic analysis was performed on founder line #295. |
Molecular Note | The human NPHS2 promoter, including the entire 5' untranslated region, was used to drive cre recombinase expression in podocytes. |
Mutations Made By | Lawrence Holzman, University of Pennsylvania |
Allele Name | targeted mutation 1.1, Christian Gachet |
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Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | MYH9Flx; Myh9flox |
Gene Symbol and Name | Myh9, myosin, heavy polypeptide 9, non-muscle |
Gene Synonym(s) | |
Strain of Origin | 129 |
Chromosome | 15 |
Molecular Note | A loxP site was inserted upstream of exon 1, and a floxed neo cassette was inserted downstream of exon 1. Transient expression in ES cells removed the neo cassette leaving exon 1 floxed. |
While maintaining a live colony, these mice are bred as double homozygotes.
When using the FVB.Cg-Myh9tm1.1Gac Tg(NPHS2-cre)295Lbh/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36742 in your Materials and Methods section.
Facility Barrier Level Descriptions
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.
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