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| Caption | Generation of the Elk3tm1.1Ics and Elk3tm1.2Ics alleles. The targeting vector was made as follows. The 5' (4.3 kb), 3' (3 kb) and inter-loxP (2.37 kb) fragments were PCR amplified on 129sv genomic DNA and sequentially subcloned in an Institut Clinique de la Souris (ICS) proprietary vector containing the LoxP sites and a Neo cassette flanked by FRT sites. The linearized construct was electroporated in 129S2/SvPas mouse embryonic stem (ES) cells. After selection, targeted clones were identified by PCR using external primers and further confirmed by Southern blotting with 5' external probe. Two positive ES clones were injected into C57BL/6J blastocysts, and derived male chimeras gave germline transmission. The excision of the neomycin-resistance cassette was performed in vivo by breeding the chimeras with a Flp deleter line (C57BL/6J genetic background) to generate the Elk3tm1.1Ics allele. The Flp transgene was segregated by breeding the first germ line mice with a wild type C57BL/6J animal. Constitutive knock-out allele, Elk3tm1.2Ics, was generated by breeding floxed-allele heterozygotes with a Cre deleter line followed by segregation in a further breeding step. The initial targeting vector contained a deletion of 104 bp (indicated by a red triangle), which was inconsequential regarding generation of the KO locus. Subseqent to Cre activation, deletion of genomic sequences extends from - 1720 to +650 relative the major transcription start site (TSS, +1), which corresponds to the first nucleotide of Elk3-001 ENSMUST00000008542 (mouse GRCm38, Ensembl). The location of primers used for genotyping is indicated by P1, P3 and P4. Use of primers P1 and P3 detect the WT allele (PCR product size 363 bp), use of primers P3 and P4 amplify sequences surrounding the region that is deleted in the KO, and therefore detect a shorter fragment in Elk3 KO mice (PCR product size 230 bp, the 2480 bp product is not amplified under the PCR conditions used). Features are not drawn to scale. | ||||||
| Copyright | This image is from Weinl C, PLoS One 2014;9(9):e107048, and is displayed under the terms of the Creative Commons Attribution 4.0 International License. J:221521 | ||||||
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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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