| Primary Identifier | MGI:5303293 | Allele Type | Targeted |
| Attribute String | Reporter | Gene | Hprt1 |
| Transmission | Cell Line | Strain of Origin | (B6.129P2-Hprt1<b-m3>/J x 129S-Gt(ROSA)26Sor<tm1Sor>/J)F1 |
| Is Recombinase | false | Is Wild Type | false |
| Project Collection | Pleiades Promoter Project |
| molecularNote | The Ple159-EGFP plasmid (pEMS1355) was created by inserting the Ple159 MiniPromoter (derived from specific promoter/enhancer/regulatory regions of the human paired-like homeodomain 3 (PITX3) gene) into the multiple cloning site of the pEMS1306 plasmid vector backbone. This placed the Ple159 MiniPromoter upstream of a minimal F5 mutant-frt site, a Kozak consensus sequence, an enhanced green fluorescent protein (EGFP; with mutated TAA->TTA stop), a tetracysteine residue, a nuclear localization signal, a minimal wildtype frt site, an SV40 early polyA signal, a human HPRT complementary sequence (containing exon 1, intron 1, exon 2, and part of intron 2), a mouse 3' Hprt homology arm, an I-Sce linearization cut site, and a mouse 5' Hprt homology arm. This Ple159-EGFP construct was targeted as a single copy knock-in to the Hprtb-m3 mutant locus on the X Chromosome via electroporation into mEMS1218 embryonic stem cells: a male ES cell line from a B6129F1-Gt(ROSA)26Sortm1Sor/+ Hprtb-m3/Y mouse (F1 generation male pups born of C57BL/6J-congenic females homozygous for a Hprtb-m3 mutation [originally derived from 129P2/OlaHsd-derived E14TG2a embryonic stem cells] X 129S-Gt(ROSA)26Sortm1Sor heterozygous males (see Stock No. 002171 and 003310, respectively)). The resulting ES cells, correctly targeted with the Ple159-EGFP transgene inserted into the Hprt locus, were assigned Hprttm207(Ple159-EGFP)Ems. |
