B6.Cg-Tg(Pax8-rtTA2S*M2)1Koes/J

Stock number: 007176
Research areas: Developmental Biology Research, Endocrine Deficiency Research, Internal/Organ Research, Research Tools

Description

These Pax8-rtTA mice provide a Tet-On tool that allows the inducible expression of genes in renal tubular epithelial cells, and may be useful in studying renal disorders such as fibrosis or polycystic kidney disease, renal cancer, and Tuberous Sclerosis (along with Stock No. 005680).

Detailed description

Transgenic Pax8-rtTA mice are viable and fertile. These mice express an optimized reverse tetracycline-controlled transactivator (rtTA2^S-M2) protein under the control of the murine Pax8 promoter, which directs expression to proximal and distal tubules and the collecting duct system of both embryonic and adult kidney. The rtTA2^S-M2 variant of rtTA contains five amino acid changes in the TetR moiety (S12G, E19G, A56P, D148E, and H179R) and a synthetic optimized transactivating domain, resulting in reduced basal activity and enhanced doxycycline sensitivity compared to wild-type rtTA. When mated to a second strain carrying a gene of interest under the regulatory control of a tetracycline-responsive promoter element (TRE or tetO), expression of the target gene in kidney cells is induced with administration of the tetracycline analog, doxycycline (dox). These Pax8-rtTA mice provide a Tet-On tool that allows the inducible expression of genes in renal tubular epithelial cells, and may be useful in studying renal disorders such as fibrosis or polycystic kidney disease, renal cancer, and Tuberous Sclerosis.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

The Pax8-rtTA construct was designed to contain an optimized rtTA variant (rtTA2s-M2) cDNA and SV40 polyA sequence replaced at the endogenous ATG translational start site of a murine Pax8 sequence. This 6.6 kb transgene was microinjected into the pronucleus of one-cell fertilized mouse embryos obtained from superovulated F2 (C57BL/6 x DBA) females, which were then implanted into pseudopregnant foster mice. Founder mice were bred to C57BL/6 mice, and the resulting transgenic offspring were bred together for many generations prior to arrival at The Jackson Laboratory. Upon arrival, some mice were backcrossed to C57BL/6J for at least 5 generations to generate this congenic strain (Stock No. 007176). The Pax8-rtTA construct integrated approximately five copies in a head-to-tail orientation within a mouse L1 line repetitive element (which are located throughout the genome).

Allele

Symbol: Tg(Pax8-rtTA2S*M2)1Koes
Name: transgene insertion 1, Robert Koesters
MGI accession ID: MGI:3709326
Generation method: Transgenic
Attributes: Transactivator
Marker symbol: Tg(Pax8-rtTA2S*M2)1Koes
Marker name: transgene insertion 1, Robert Koesters
Chromosome: 8
Strain of origin: (C57BL/6 x DBA)F2
Expressed genes: rtTA,reverse tetracycline-controlled transactivator,E. coli
Site of expression: expression is directed to proximal and distal tubules and the collecting duct system of both embryonic and adult kidney.
Molecular note: The Pax8-rtTA construct contains an optimized rtTA variant (rtTA2s-M2) cDNA and SV40 polyA sequence replaced at the endogenous ATG translational start site of a murine Pax8 sequence. The rtTA2S*M2 variant contains 5 amino acid changes in the TetR moiety (S12G, E19G, A56P, D148E, and H179R) and a synthetic optimized transactivating domain, resulting in reduced basal activity and enhanced doxycycline sensitivity compared to wild-type rtTA.