| Primary Identifier | MGI:7860774 | Allele Type | Targeted |
| Attribute String | Conditional ready, Inducible, Null/knockout | Gene | Dnmt1 |
| Transmission | Germline | Strain of Origin | C57BL/6 |
| Induced With | doxycycline/tetracycline | Is Recombinase | false |
| Is Wild Type | false |
| molecularNote | F.A.S.T. (Flexible Accelerated STOP TetO) cassette consisting of a loxP-Frt-neo-Frt-STOP-Frt-PTRE3G-loxP sequence and Not I cloning site, knocked into the Dnmt1 gene locus. A 9.6 kb region was subcloned from a positively identified C57BL/6 fosmid clone (WI1-560618) and used to assemble a targeting vector carrying the F.A.S.T. cassette, consisting of a loxP-Frt-neo-Frt-STOP-Frt-PTRE3G-loxP sequence and a Not I cloning site. A Not I site downstream of the exon 1 of Dnmt1 was modified to allow the cloning of the F.A.S.T. knock-in cassette. The short homology arm (SA) of 2.8 kb and the long homology arm (LA) of 6.7kb enable the homologous recombination. The targeting vector was confirmed by restriction analysis and sequencing after each modification step. The boundaries of the vector were confirmed by sequencing with primers P6 and T73. The F.A.S.T. cassette sequence was confirmed by sequencing with primers SQ1, P2, and FAST SQ1. The point mutation to removes the Not I site was confirmed by FAST SQ1. |
