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Title: Clinical and experimental studies on the role of APC in neoplasia
Author: Cooper, Cindy Anne
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2001
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This thesis describes firstly, the generation and initial characterisation of a murine APC transgenic founder line designed as a model to investigate the effects of aberrant expression of APC. Several Apc mutant murine models of FAP already exist. These all carry a heterozygous mutation in the Apc gene. The Min mouse (multiple intestinal neoplasia) is an example and carries a mutation at codon 850. All current models however are of limited use as mice homozygous for Apc mutations die at approximately 6.6 days post coitum, limiting analysis to Apc heterozygotes. Homozygous loss of Apc therefore depends upon additional somatic events that are not under direct experimental control and this may be associated with additional, undisclosed genetic events. Here a transgenic approach has been taken to generate animals where the expression of the APC transgene is conditionally inactivated using the Cre-lox P recombination system of the bacteriophage P1. APC cDNA was flanked by lox P sequences and the phosphoglycerate kinase (Pgk) ubiquitous promoter DNA sequence cloned upstream of the APC cDNA to drive the expression of the gene. Pro-nuclear injection was used to deliver the APC transgene into oocytes of wild type mice (F1 (C57B1/6 x CBA)). In one founder line transgene copy number was determined by southern blotting and transcription of the APC cDNA confirmed in a wide variety of tissues using reverse transcription polymerase chain reaction (PCR). Transgenic positive mice on an Apc wildtype background showed no gross phenotype. Embryonic fibroblasts were derived and Cre-recombinase delivered by infection with a replication deficient adenovirus. APC cDNA excision was confirmed by PCR. The founder line was crossed with the C57BL/6 Apc Min/Wt line. Transgenic positive Apc Min/Wt mice were intercrossed and offspring screened to identify whether the APC transgene can rescue the Apc Min/Min lethal phenotype. To date embryonic rescue has not been identified.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available