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| Caption | Generation of the Trex1tm1.1Fwpe allele. Replacement of the TREX1 wild-type (WT) allele with the TREX1 D18N allele. The R1 129 mouse TREX1 genomic region (~10kb) flanked by SacI sites (lower) and the vector used for targeted homologous recombination (upper) are shown (Boxes, exons; lines, introns; squiggle lines, mouse genomic sequences flanking the region of homology; D18N codon, point mutation introduced into the vector TREX1 coding exon and C->T change in TREX1 5' UTR; NEO, neomycin resistance gene; DT-A-, diphtheria toxin A gene; LoxP, recognition site for Cre recombinase; PCR1-7, PCR primers). The linearized TREX1 D18N vector was electroporated into XSV1 ES cells (of a 129S6/SvEvTac origin) and DT-A G418-resistant colonies were selected for screening using PCR1/PCR2 and PCR3/PCR4 to identify candidate ES cell clones for targeted homologous recombination. Both PCR3 and PCR4 contain a 3' terminal mispair unless the ES clone contains the targeted mutant allele. PCR positive clones for both reactions with the expected size PCR products had integrated the NEO linked vector into the TREX1 locus. A third PCR using PCR5/ PCR6 recovers a 1.4 kb genomic fragment and sequencing confirmed targeting of the Trex1 mutant allele and the appropriate heterozygosity corresponding to the presence of both the WT and mutant alleles in the ES cell clones. Cre-mediated recombination removed the NEO cassette. | ||||
| Copyright | This image is from Grieves JL, Proc Natl Acad Sci U S A 2015 Apr 21;112(16):5117-22. Copyright 2015 National Academy of Sciences, U.S.A. J:220965 | ||||
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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 09/03/2024 MGI 6.24 |
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