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| Caption | Conditional Ser133Ala knockin in Creb1. (a) Exon usage of the three main CREB splicing isoforms, CREBa, D, and b. Ser133 is encoded in Exon 5 of CREBa, and is present in all three isoforms. (b) The targeting vector to generate the conditional Creb1 allele was designed to insert a minigene and neomycin (neo) resistance gene upstream of Exon 5. The minigene consisted of a cDNA corresponding to exons 5-9 fused to the splice acceptor site of Exon 5. LoxP sites (red triangles) were placed before the minigene and after the neomycin gene. Frt sites (blue triangles) were placed either side of the neomycin gene. The copy of Exon 5 in the 30 arm of homology was mutated to change the codon for Ser133 to alanine (indicated by *). Endogenous Bam HI and Kpn I sites are indicated by black arrows, additional sites introduced during PCR cloning of the arm to facilitate cloning or screening are indicated by grey arrows. (c) In the allele containing the minigene transcription of the endogenous promoter should result in splicing from Exon 4 onto the minigene, followed by transcriptional termination at the end of the minigene. This would result in the expression of wild type CREB. Expression of Cre recombinase would result in excision of the minigene. Splicing would then occur from Exon 4 onto the mutated copy of Exon 5, resulting in the expression of the Ser133Ala knockin mutation. | ||||
| Copyright | This image is from Wingate AD, Genesis 2009 Jul 20;47(10):688-696, and is displayed with the permission of Wiley-Blackwell, who owns the Copyright. J:153559 | ||||
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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 10/09/2024 MGI 6.24 |
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