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| Caption | (Top) Targeting vectors (TV) for the H3f3btm1.1Mnn (conditional H3f3b (c) allele) and H3f3btm1.2Mnn (mutant H3f3b (-) allele) alleles. The core region of cassettes was nearly identical for H3f3a and H3f3b except that the 3' UTR was specific for the gene. This core sequence substituted for much of exon 2 for H3f3a and all of exon 2 for H3f3b, encoding the first 17 and 41 aa of H3.3, respectively. Exposure to Flp recombinase removed the neo selection cassette to produce the final conditional allele. The cassette arrangement and positioning of the loxP sites allows conditional allelic replacement: on Cre-mediated excision of the H3.3 coding sequence (cds), an alternative cds is brought under control of the promoter, this being the yfp cds for H3f3a, and the cfp cds for H3f3b. These TVs have been designed such that a new proxy cds can be inserted in place of theyfp or cfp cds in one cloning step. In these newer vectors, the 3' UTR has already been inserted such that it will be downstream of the proxy. The horizontal arrow indicates progression of the transcript from the promoter (Prom). The yellow shading marks the regions where strand exchange would be expected to occur. The red line marks a potential and undesired internal strand exchange (ISE) event occurring due to internal regions of homology. Such events would lead to loss of upstream TV sequence including the loxP site. SpA, synthetic splice acceptor; ex, exon; neo, neomycin positive selection cassette; dtA, diphtheria toxin A chain negative selection cassette; E, EcoRI, K, KpnI; N, NheI site; X, XhoI; pBS, pBluescript vector. (Bottom) Schematic of the H3f3atm1.1Mnn (H3f3b conditional allele) and H3f3btm1.2Mnn (H3f3b mutant allele) alleles. The horizontal arrow and dotted line indicates progression of the transcript from the promoter (Prom). The black vertical arrow indicates probable foreign poly(A) site usage. The value above the arrow is the predicted transcript size without taking into account additional sequence added as poly(A) tract. Gray vertical arrow indicates the probable endogenous poly(A) site used. | ||||||
| Copyright | This image is from Tang MC, Genesis 2013 Feb;51(2):142-6, and is displayed with the permission of Wiley-Blackwell, who owns the Copyright. J:194069 | ||||||
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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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