These Nephrin KO/GFP knock-in mice may be useful in studying nephrotic syndrome or any of the diseases where plasma ultrafiltration is affected.
Raghu Kalluri, University of Texas MD Anderson Cancer Center
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Reporter, Null/Knockout) | Nphs1 | nephrosis 1, nephrin |
Mice that are heterozygous for the Nephrin KO/GFP knock-in mutation are viable and fertile. Homozygous mice die within a few days after birth with massive glomerular vascular leak and an absence of glomerular epithelial slit diaphragms. No gene product is detected by Western blot of kidney tissue of homozygotes. Green fluorescent protein (GFP) is detected in glomeruli of homozygotes and heterozygotes. Newborn homozygotes show extensive proteinuria, while heterozygotes show none. These Nephrin KO/GFP knock-in mice may be useful in studying nephrotic syndrome or any of the diseases where plasma ultrafiltration is affected.
A targeting vector was generated in which base pairs -1 to 33 surrounding the ATG start codon of exon 1 of the endogenous gene were replaced with an in-frame green fluorescent protein-polyadenylation stop followed by a phosphoglycerol kinase promoter-neomycin resistance-polyadenylation stop. The construct was electroporated into 129/Sv derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and the resulting chimeric males were bred to 129/Sv females. The donating investigator reports that heterozygotes were backcrossed to 129/Sv for more than ten generations. Upon arrival at The Jackson Laboratory, mutant mice were bred with 129S1/SvImJ inbred mice (Stock No. 002448) for at least one generation to establish the colony.
A 27 SNP (single nucleotide polymorphism) panel analysis performed by The Jackson Laboratory revealed 9 of 27 markers (on nine different chromosomes) that were not 129 allele-type. These data suggest C57BL/6 (and perhaps other) genetic contamination prior to arrival at The Jackson Laboratory.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Site of Expression | GFP is detected in glomeruli. |
Allele Name | targeted mutation 1, Raghu Kalluri |
---|---|
Allele Type | Targeted (Reporter, Null/Knockout) |
Allele Synonym(s) | |
Gene Symbol and Name | Nphs1, nephrosis 1, nephrin |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | GFP is detected in glomeruli. |
Strain of Origin | 129/Sv |
Chromosome | 7 |
General Note | Immunohistochemical and western blot analyses of kidneys of homozygous newborn mice demonstrated absence of nephrin protein. No alteration was observed in expression of any glomerular basement membrane (GBM) or glomerular epithelial slit diaphragm (GESD) protein other than nephrin; proteins analyzed included integrin alpha3, podocin, alpha actinin 4, CD2AP, synaptopodin, nephronectin and type IV collagens alpha3, alpha4 and alpha5. (J:78616) |
Molecular Note | 34 bp surrounding the translational start codon in exon 1 (-1 through +33) were replaced with a GFP-neo cassette inserted via homologous recombination. Expression of GFP was observed in both homozygous and heterozygous mutant mice. |
Mutations Made By | Raghu Kalluri, University of Texas MD Anderson Cancer Center |
When maintaining a live colony, heterozygous mice may be bred together or to wildtype siblings. Homozygous mice a few days after birth.
When using the Nephrin KO/GFP knock-in mice mouse strain in a publication, please cite the originating article(s) and include JAX stock #005692 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Nphs1<tm1Rkl> |
Frozen Mouse Embryo | STOCK Nphs1<tm1Rkl>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Nphs1<tm1Rkl>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Nphs1<tm1Rkl>/J | $3373.50 |
Frozen Mouse Embryo | STOCK Nphs1<tm1Rkl>/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.