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Analysis of meiotic, apoptosis-inducing checkpoint pathways by genomics-based, forward genetic and two hybrid approaches in C. elegans

Subject Area General Genetics and Functional Genome Biology
Term from 2001 to 2005
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 5339398
 
The main goal of this proposal is to use the C. elegans experimental system to define evolutionarily conserved meiotic checkpoint pathways that sense mistakes occuring during the meiotic prophase, like meiotic chromosome pairing and meiotic recombiantion defects and trigger the apoptotic demise of affected cells, by 'functional genomics' based techniques. 1) We postulate that components of meiotic checkpoint pathways are specifically transcibed in meiotic cells. We will inactivate all, approximately 750 genes that are enriched in the germ line by RNAi feeding. Double stranded RNA (RNAi) against a gene of choice applied to a worm leads to functional gene inactivation. RNAi is most conveniently applied to worms by feeding them with E. coli expressing dsRNA. To further our understanding of meiotic checkpoint/apoptosis signaling we well screen for the suppresion of germ cell apoptosis that is induced by meiotic recombination and chromosome pairing defects. In addition to the forward genetic, RNAi-based screen, we will 2) analyze whether known conserved DNA damage response genes also affect meiotic checkpoint regulation. Furthermore, 3) two hybrid interactors of these checkpoint proteins will be assessed for specific meiotic checkpoint functions. 4) The molecular biology of a selected set of conserved and novel meiotic checkpoint proteins will be analyzed by genetic, biochemical and cytological techniques.
DFG Programme Research Grants
 
 

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