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Functional analysis of regulated clathrin heavy chain depletion in a vertebrate cell line

Obiettivo



Research objectives and content
We are exploiting the high rate of homologous recombination in the chicken pre-B cell line DT40 toinactivate the clathrin heavy chain gene (CHc) by gene targeting. We have generated CHc-heterozygotesbut failed to produce cells lacking both functional alleles, suggesting a lethal phenotype. To circumventthis problem we established cells expressing human CHc under the control of the inducible Tet-system.This will enable us to generate a CHc-null phenotype and investigate the functional consequences ofregulated clathrin depletion. The project aims to understand why the disruption of the CHc gene is lethal in DT40. The Tet-system will enable us to study the response of clathrin-mediated pathways to restricted CHc-levels and to investigate possible alternative pathways. Site-directed mutagenesis will gain new insight into the mechanism of clathrin-coated vesicle formation.
Training content (objective, benefit and expected impact)
Dr Jackson's group has experience in molecular and cell-biological aspects of clathrin function. Theculmination of this work, in a laboratory with expertise in the field of coated vesicle formation, will be the pursuit of our system to the point where functional studies can be conducted. In addition to the specificfacilities the biochemistry department can offer (see detailed proposal), Cambridge is also an excellent centre for related questions as there are several groups that have similar interests and that have aninternational reputation. Links with industry / industrial relevance (22)
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Campo scientifico (EuroSciVoc)

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Meccanismo di finanziamento

RGI - Research grants (individual fellowships)

Coordinatore

THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
Contributo UE
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Indirizzo
80,Tennis Court Road, 80
CB2 1GA CAMBRIDGE
Regno Unito
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