| Primary Identifier | MGI:5303267 | 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 RP11-406M16 bacterial artificial chromosome (BAC) was obtained and retrofitted to contain 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; all inserted into the sacB gene in the BAC vector backbone. Next, an enhanced green fluorescent protein (EGFP) and frt-flanked kanamycin cassette were inserted directly at the ATG of the human lactase (LCT) gene; this created the Ple126-EGFP BAC (bEMS75, LCT). The kanamycin cassette was transiently removed via FLP recombination; leaving the EGFP gene followed by a single frt site. The final modified BAC was used as the source for the Ple126-EGFP transgene. The Ple126-EGFP transgene was targeted as a single copy knock-in to the Hprtb-m3 mutant locus on the X Chromosome via electroporation into mEMS1204 embryonic stem cells. |
