| Primary Identifier | MGI:7860772 | Allele Type | Targeted |
| Attribute String | Transactivator | Gene | Tcf21 |
| Transmission | Germline | Strain of Origin | C57BL/6 |
| Is Recombinase | false | Is Wild Type | false |
| molecularNote | Reverse Tet-off rtTA protein controlled by Tcf21 promoter (fibroblast-specific). A 9.1 kb region was first subcloned from a positively identified C57BL/6 fosmid clone (WI1-336E22) and used to build the targeting vector using homologous recombination-based techniques. The region is designed so that the long homology arm (LA) extends 6.2 kb upstream of the P2A-rtTA knock-in cassette and the short homology arm (SA) extends 2.3 kb downstream of the neo cassette. The P2A-rtTA cassette was inserted upstream of the TGA stop codon in exon 2 of the mouse Tcf21 gene. The FRT flanked neo selection cassette followed downstream of the 3â-UTR sequence. The targeting vector was confirmed by restriction analysis and sequencing after each modification step. The boundaries of the 2 homology arms were confirmed by sequencing with primers P6 and T73 that read through both sides of the backbone vector into the genomic sequences. The P2A-rtTA knock-in cassette was confirmed by sequencing with primers TCFC SQ1 and SQ2. IV-neo N3 and ineo N2 primers, reading from the selection cassette, confirmed the insertion of the neo cassette. The PTRE3G promoter has been used. |
