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| Caption | Conditional Cre knock-in construct at the Pou4f2 (Brn3b) locus generating the Pou4f2tm1.1(cre)Tcba allele. Diagrams represent the targeting strategy and recombination steps for the Pou4f2 (G-J) locus. J, wild-type configuration, G, locus after gene targeting, H, locus after removal of the Neo selection cassette by cross with germline Flp recombinase expressing mice which results in the Pou4f2tm1.1(cre)Tcba allele, I, 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 Pou4f2 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 (I) 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 Pou4f2, Kpn I digests are tested with the 5' probe, recognizing fragments of 12624 bp in wild-type, and 14388 bp in the targeted construct, before and after Neo cassette removal; Bam HI digests, tested with the 3' probe yielding characteristic fragments of 10140 bp in the wild type, 6772 bp in the targeted locus, and 7682 bp after Neo 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 11/12/2024 MGI 6.24 |
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