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| Caption | Conditional Cre knock-in construct at the Pou4f1 (Brn3a) locus generating the Pou4f1tm1.1(cre)Tcba allele. Diagram represents the targeting strategy and recombination steps for the Pou4f1 (A-D) locus. D, wild-type configuration, A, locus after gene targeting, B, locus after removal of the Neo selection cassette by cross with germline Flp recombinase expressing mice which results in the Pou4f1tm1.1(cre)Tcba allele, C, locus after removal of the endogenous gene by Dre mediated recombination, resulting in expression of the Cre recombinase gene from the original transcription start site of the Pou4f1 gene. Tick marks represent 1 kb distances, except for contracted longer stretches marked with SS marks. Green and white boxes represent coding regions and UTRs respectively, and splice junctions are indicated by angled black lines. Homologous recombination targeting arms are indicated by black lines, and southern blot probes as black boxes under the locus. Diagnostic restriction enzymes are represented as capital letters: Nde I (N), Kpn I (K), and Bam HI (B). A triple repeat of the SV40 polyA is inserted in the 3' UTR of the targeted locus, and indicated by a red thick arrow. RoxP sites (white circles), flank the endogenous gene while FRT sites (blue circles), flank the PGK-Neo Cassette. Genotyping primers (thin black arrowheads) are placed in the 5' UTR of the loci, and encompass the ATG in the wild-type and ATG and 5' roxP site in the targeted locus. After Dre recombination (C) the reverse strand black primer is lost, and correct localization of the Cre can be tested by the black and white pair of primers indicated. Southern blot fragment lengths are as follows: for Pou4f1, the Nde I fragment is recognized by both 5' and 3' probes, and is 19116 bp for the wild-type, 23542 for the targeted locus, and 21637 bp for the targeted locus after Flp mediated Neo cassette removal. | ||||
| Copyright | This image is from Sajgo S, PLoS One 2014;9(3):e91435, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:215125 | ||||
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Associated Alleles |
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Mouse Genome Database (MGD), Gene Expression Database (GXD), Mouse Models of Human Cancer database (MMHCdb) (formerly Mouse Tumor Biology (MTB)), Gene Ontology (GO) |
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last database update 05/07/2024 MGI 6.23 |
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