Mouse Strains: Id3
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Id3 expression is not confined to a known cell type within the developing metanephros. An Id3 knock-in approach was employed by the GUDMAP consortium as a means of investigating this population of cells in more detail.
A transgenic founder is mated to C57Bl6 to establish the line and provide E15.5 urogenital systems for analysis. Identification of the transgenic animals is based on direct detection of dsRED fluorescence, marking gene expression in the dissected urogenital system. Transgenic animals are confirmed by PCR genotyping.
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| Gross dissection of E15.5 urogenital systems and RFP visualisation. |
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| Native RFP fluorescence viewed at E15.5. Though the cortex is poorly detected in this image, the observed pattern of RFP agrees with the in situ pattern documented (GUDMAP:7218). |
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| Confocal imaging of E15.5 sections showing appropriate RFP expression in the Id3 expression domain. The Dapi counterstain is false coloured green here for contrast to the expression in red. The last row shows a littermate control at the same exposure settings. |
PCR primer sequences used to genotype trangenic animals are provided below:
| Id3-F | 5’ tcc tcg gta tca gcg ctt cc 3’ |
| Id3-R | 5’ caa tgg cca ggc tac gtt cc 3’ |
| RFP-R | 5’ ctt gat gac gtc ctc gga gg 3’ |
The expected band sizes of these PCR products are as follows:
| Id3-F + Id3-R | 351 bp |
| Id3-F + RFP-R | 234 bp |
Relevant Sequences
Click on the following links to access sequence data relating to the targeting construct:
| Genomic clone | Final construct | |
| (Genomic clone used for targeting construct) | (excludes plasmid backbone and the negative selection marker) |