STOCK Tg(Hnf1b-cre/ERT2)1Jfer/J

Stock number: 027681
Product name: Hnf1bCreER
Research areas: Endocrine Deficiency Research, Research Tools

Description

Hnf1bCreER mice may be useful for generating specific targeted mutations or fate-mapping studies of Hnf1b expressing cells.

Detailed description

Hnf1bCreER mice express Cre-ER^T2 directed by the murine Hnf1b (HNF1 homeobox B) promoter/enhancer regions. The construct also contains an internal ribosomal recognition site (IRES) and a destabilized enhanced green fluorescent protein (D2EGFP), however the donating investigator reports that the D2EGFP is not detected by endogenous fluorescence or immunofluorescence. These mice express Cre-ER^T2 protein in all of the cellular types where Hnf1b is highly expressed, such as pancreatic duct epithelium, hepatic ductal epithelium, kidney, and gut. Restricted to the cytoplasm, Cre-ER^T2 can only gain access to the nuclear compartment after exposure to tamoxifen. Mice homozygous for the transgene are viable and fertile. When these mice are bred with mice containing loxP-flanked sequence, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.

Development

Bacterial artificial chromosome (BAC) library (CHORI) was used to obtain a BAC (#RP23-304H7) containing the entire mouse Hnf1b (HNF1 homeobox B) gene. This BAC was modified to have exon 1 replaced with a creERT2 fusion gene (a Cre recombinase fused to a human estrogen receptor ligand binding domain), an internal ribosomal recognition site (IRES), a destabilized enhanced green fluorescent protein (D2EGFP), and a polyadenylation sequence. The BAC was microinjected into fertilized B6SJL F1 oocytes and founder mice, carrying one copy of the transgene, were identified. Subsequently the Hnf1bCreER mice were crossed with other strains and were maintained on a mixed background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Tg(Hnf1b-cre/ERT2)1Jfer
Name: transgene insertion 1, Jorge Ferrer
MGI accession ID: MGI:4420452
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Marker symbol: Tg(Hnf1b-cre/ERT2)1Jfer
Marker name: transgene insertion 1, Jorge Ferrer
Chromosome: UN
Strain of origin: C57BL/6J x SJL
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: Cre-ERT2 fusion protein is expressed in Hnf1b -expressing cells in pancreatic duct epithelium, hepatic ductal epithelium, kidney, and gut.
Molecular note: A BAC clone containing the Hnf1b locus (RP23-304H7) was modified by replacing exon 1 with a targeting vector containing an Hnf1b promoter fragment, cre/ERT2, IRES, d2EGFP (a destabilized variant of EGFP), the b-globin polyadenylation site, a FRT/neo/FRT cassette, and an intron 1 fragment that flanks exon 1 of Hnf1b. The targeting vector was electroporated into EL250 cells and homologous recombination was induced. Correctly targeted clones were treated to activate inducible Flp recombinase, removing the FRT/neo/FRT cassette. The recombined BAC was digested, linearized and microinjected into pronuclei of C57BL6/SJL eggs. 10 lines were generated, and after crossing to Rosa26 reporter mice, one line carrying a single copy of the transgene was maintained (after characterization of 4 lines exhibiting similar reporter recombination patterns.)