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Caption (A) Scheme depicting the generation of the Gt(ROSA)26Sortm1Npa allele in ES cells. A Stop sequence and a HygR selection cassette flanked by heterospecific LoxP sites (Lox511 and LoxP) were targeted to the Rosa26 locus between exons 1 and 2 by homologous recombination. After successful recombination, the Stop cassette is located downstream of the endogenous Rosa26 promoter. (B) Cre-RMCE into the Gt(ROSA)26Sortm1Npa (modRosa26LoxP) locus. In the Cre-RMCE targeting plasmid, a promoter, EGFP and an FRT-flanked neomycin selection cassette (NeoR) were flanked by heterospecific LoxP sites (Lox511 and LoxP) as a group. Cre-RMCE was used to replace the HygR in the mutant ES cells with the Lox511/LoxP-flanked sequence in the RMCE targeting plasmid. A pCAG, CMV or EEF1a1 (EF1a) promoter driving EGFP was introduced resulting in the Gt(ROSA)26Sortm2(CAG-EGFP)Npa, Gt(ROSA)26Sortm3(CMV-EGFP)Npa, and Gt(ROSA)26Sortm4(EEF1A1-EGFP)Npa alleles, respectively. Insertion of EGFP without any promoter (NoP) controls for functional shielding of the integration site from the endogenous Rosa26 promoter. Binding regions for TaqMan genotyping primers (1, 3) and probe (2), primers for checking integration into the modRosa26LoxP locus (4-7) and the Southern hybridization probe, as well as the BamHI sites used for Southern blot analysis are indicated.
Copyright This image is from Tchorz JS, PLoS One 2012;7(1):e30011, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:184312
Associated
Alleles
Symbol Name
Gt(ROSA)26Sortm2(CAG-EGFP)Npa gene trap ROSA 26, Philippe Soriano; targeted mutation 2, Novartis Pharma AG
Gt(ROSA)26Sortm3(CMV-EGFP)Npa gene trap ROSA 26, Philippe Soriano; targeted mutation 3, Novartis Pharma AG
Gt(ROSA)26Sortm4(EEF1A1-EGFP)Npa gene trap ROSA 26, Philippe Soriano; targeted mutation 4, Novartis Pharma AG

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last database update
11/19/2024
MGI 6.24
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