B6.Cg-Tg(NPHS2-Trpc6)F419Walz/J

Stock number: 018293
Product name: Trpc6-HA wt
Strain synonyms: pNPHS2 Trpc6-HA wt F419 transgenic mice
Research areas: Developmental Biology Research, Endocrine Deficiency Research, Internal/Organ Research

Description

Hemizygous "pNPHS2 Trpc6-HA wt F419" transgenic mice have ~4-fold overexpression of mouse wildtype Trpc6 (with 3' hemaglutinin tag) directed to glomerular podocytes by the human podocin promoter/enhancer sequences (NPHS2). These transgenic mice may be useful for studying kidney disease of the glomerulus (including albuminuria, podoyte structural injuries, glomerular lesions and tubule-interstitial damage), as well as pathological features of Focal and Segmental Glomerulosclerosis (FSGS).

Detailed description

Hemizygous pNPHS2 Trpc6-HA wt F419 transgenic mice are viable and fertile, with overexpression of mouse wildtype Trpc6 (with 3' hemaglutinin tag) directed to glomerular podocytes by the promoter/enhancer region of the human podocin locus (NPHS2). The donating investigator reports mice from founder line F419 have ~2 copies of the transgene and, using glomerular samples purified by dynabead extraction, glomerular overexpression levels are ~4 times greater than endogenous Trpc6 mRNA. Expression of the transgene is restricted to glomerular podocytes (as illustrated by a strong co-localization with the podocyte-specific marker synaptopodin). The HA-tag functions for use in hemaglutinin immunoprecipitation. 10-30% of transgenic males between 5-9 months of age exhibit albuminuria, podoyte structural injuries, glomerular lesions, tubule-interstitial damage, and pathological features of Focal and Segmental Glomerulosclerosis (FSGS). Electron microscopy reveals ultrastructural podocyte changes as early as ~3 months of age in transgenic mice.

Development

The Trpc6-HA wildtype cDNA sequence was made by modifying a Trpc6 cDNA sequence (encoding a 930 amino acid protein) originally isolated from mouse brain tissue by both repairing an A₂₆₁₅G point mutation (D872G) and also by in-frame addition of an influenza virus hemaglutinin epitope (HA tag) at the 3' end. The cDNA sequence was placed downstream of a 2.5 kbp fragment from the promoter/enhancer region of the human podocin locus (NPHS2) and upstream from a bovine growth hormone polyA sequence. The resulting "pNPHS2 Trpc6-HA wt" transgene was microinjected into the pronuclei of (C57B6/6J x CBA/J) zygotes. Founder mice were bred with C57BL/6J mice to establish line F419. At the F2 generation, mice from founder line F419 had ~2 copies of the transgene. Transgenic mice were subsequently maintained by backcrossing to C57BL/6J for five generations (~96.9% C57BL/6J) prior to sending to The Jackson Laboratory Repository. Upon arrival, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish The Jackson Laboratory Repository colony.

Allele

Symbol: Tg(NPHS2-Trpc6)F419Walz
Name: transgene insertion F419, Katherina Walz
MGI accession ID: MGI:5319631
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(NPHS2-Trpc6)F419Walz
Marker name: transgene insertion F419, Katherina Walz
Chromosome: UN
Strain of origin: C57BL/6J x CBA/J
Expressed genes: Trpc6,transient receptor potential cation channel, subfamily C, member 6,mouse, laboratory
Molecular note: The Trpc6-HA wildtype cDNA sequence was made by modifying a Trpc6 cDNA sequence (encoding a 930 amino acid protein) originally isolated from mouse brain tissue by both repairing an A2615G point mutation (D872G) and also by in-frame addition of an influenza virus hemaglutinin epitope (HA-tag) at the 3' end. The cDNA sequence was placed downstream of a 2.5 kbp fragment from the promoter/enhancer region of the human podocin locus (NPHS2) and upstream from a bovine growth hormone polyA sequence. Line F419 contains 2-4 copies of the transgene.