| Primary Identifier | MGI:5303268 | Allele Type | Targeted |
| 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 RP11-158L10 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, a beta-galactosidase (lacZ) and frt-flanked kanamycin cassette were inserted directly at the ATG of the human nerve growth factor receptor (NGFR) gene; this created the Ple133-lacZ BAC (bEMS88, NGFR). The kanamycin cassette was transiently removed via FLP recombination; leaving the lacZ gene followed by a single frt site. The final modified BAC was used as the source for the Ple133-lacZ transgene. The Ple133-lacZ transgene was targeted as a single copy knock-in to the Hprtb-m3 mutant locus on the X Chromosome via electroporation into mEMS1202 embryonic stem cells. |
