These knock-in mice express a transactivator protein (tTA) from the endogenous promoter/enhancer elements of the Pdx1 gene in the pancreas.
Raymond MacDonald, UTSW Medical Center
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Transactivator) | Pdx1 | pancreatic and duodenal homeobox 1 |
Mice homozygous for the targeted mutation fail to develop a pancreas. Heterozygous mice have normal pancreas development but partially impaired glucose tolerance in adulthood. The substitution of the targeted gene with tTAoff inactivates the endogenous allele and places tTAoff expression under the control of the endogenous transcriptional regulatory sequences. tTA expression from the modified locus is identical to that of the normal allele; tTA mRNA is detectable in the pancreas but not in other visceral organs or salivary glands. This mutant may be useful in studies of pancreatic endocrine/exocrine function and diabetes. Heterozygotes also can be bred with transgenic mice that express a target gene under the regulation of a tetracycline-responsive element (TRE; tetO) for pancreas-specific applications.
This mutant was originally designed to be mated with mutant mice with a TRE-controlled transgene coding for the endogenous Pdx1 (Ipf1) gene along with a reporter (EGFP or beta-galactosidase gene, (see Stock No. 005699 and 005728 respectively).
A targeting vector containing a tetracycline-responsive tet-repressor/VP16 fusion transactivator (tTAoff) and the neomycin-resistance gene flanked with 4.5-kb upstream and 1.3-kb downstream homology domains was used to replace the entire coding region (both exons and the intron) of the endogenous Pdx1 locus. The construct was electroporated into the (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric males were bred with C57BL/6 females to generate heterozygotes. At some point, mutant mice were bred to B6SJL hybrid transgenic mice. The transgene was selected against in subsequent breedings, and strain is now maintained as heterozygous for the Pdx1 targeted mutation.
Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli |
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Site of Expression | Expresses tTA in pancreas. |
Allele Name | targeted mutation 1, Raymond J MacDonald |
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Allele Type | Targeted (Transactivator) |
Allele Synonym(s) | Pdx1tTA |
Gene Symbol and Name | Pdx1, pancreatic and duodenal homeobox 1 |
Gene Synonym(s) | |
Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli |
Site of Expression | Expresses tTA in pancreas. |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 5 |
Molecular Note | Exons 1 and 2, which encompass the entire coding region, were replaced with a cassette containing a tetracycline transactivator (tTA) and a neo resistance gene. RT-PCR showed tTA expression in adult pancreas tissue but not in other visceral organs or salivary glands. |
Mutations Made By | Raymond MacDonald, UTSW Medical Center |
When maintaining a live colony, these mice are maintained as heterozygotes.
When using the Pdx1tTA KI mouse strain in a publication, please cite the originating article(s) and include JAX stock #005701 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Pdx1<tm1Macd> |
Frozen Mouse Embryo | STOCK Pdx1<tm1Macd>/J Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | STOCK Pdx1<tm1Macd>/J Frozen Embryos | $2595.00 |
Frozen Mouse Embryo | STOCK Pdx1<tm1Macd>/J Frozen Embryos | $3373.50 |
Frozen Mouse Embryo | STOCK Pdx1<tm1Macd>/J Frozen Embryos | $3373.50 |
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