Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.660293
Title: Characterisation of gene-trap integrations expressed during mouse heart development
Author: Pall, Gurman Singh
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1998
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Abstract:
A gene-trap strategy in embryonic stem (ES)cells has been employed to identify and characterise genes involved in heart development. This work describes the characterisation of two gene-trap integrations (R68 and R124), including identification of endogenous trapped gene sequences and analysis to determine the function of the trapped genes. Molecular analysis of the R68 and R124 gene-trap integrations has shown the use of cryptic splice sites within the gene-trap vector indicating the vector has integrated into an exon in both gene-trap cell lines. The unpredicted integration of a gene-trap vector into an exon still results in the expression of the reporter gene. Sequence data indicates a novel gene has been trapped by the R124 gene-trap integration. The structure of the R124 integration in the genome has been predicted based upon sequencing data and restriction fragment length polymorphism analysis. The R124 gene-trap integration has been mapped to chromosome 5 in the mouse genome. Beating cardiomyoctyes generated from the in vitro differentiation of R68 and R124 ES cells express the reporter gene. Embryos heterozygous for the R124 gene-trap integration express reporter gene activity in the developing heart throughout gestation. In the adult the reporter gene is expressed in the heart, kidney, testis, ovary and brain. Function of the trapped gene was assessed by generating animals homozygous for the R124 integration. 60% of animals homozygous for the integration die shortly after birth. This lethality is associated with a right ventricle heart defect. Surviving homozygote males show enlarged hearts and kidneys. The surviving homozygote males are also infertile, histological analysis has shown no mature sperm in the testes of homozygote males.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.660293  DOI: Not available
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