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Title: Gene expression in early haematopoietic development
Author: Menzel, Ursula Rosa
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1998
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Under appropriate culture conditions, ES cells will spontaneously differentiate in vitro into a range of embryonic lineages, including the haematopoietic lineage. All haematopoietic lineages can result from ES cell differentiation including transplantable HSCs. The relative ease of transgenesis in ES cells has been exploited for the analysis of gene function and the identification of novel genes, by the use of gene targeting and gene trapping methodology, respectively. Conventional analysis of mutated ES cells is based on the production of chimeric embryos for in vivo studies. However, in vitro differentiation of mutated ES cells can provide an alternative and complementary approach to in vivo analysis. In particular an in vitro strategy for the screening of ES cells gene trap libraries could restrict the number of gene trap clones to be analyzed subsequently in chimeric embryos in vivo. Based on an established ES cell system for in vitro haematopoiesis, part of the present project has been the assessment of an in vitro prescreening strategy of a gene trap library for the identification of genes that may be involved in early haematopoietic differentiation. This was achieved by monitoring the temporal expression of a β-gal reporter gene in established gene trap lines after induction of haematopoietic differentiation by a morphogenic factor. The gene trap cell lines for use in this study were selected on the basis of their spatial expression patterns in chimeric embryos. The potential application of this strategy on a large scale has been tested by simplification of the culture procedures that support haematopoietic differentiation. The interaction of gene trap constructs into the ES cell genome facilitates the identification of the trapped endogenous gene but also allows the use of in situ hybridization for the analysis of reporter gene expression.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available