Objective
Gene targeting is of fundamental importance for basic research and in plant breeding. Since gene targeting allows directed integration of DNA into the genome a broad spectrum of applications results: The method is a valuable tool in function search in mammalian cells and will be especially useful in plants for defining functions for DNA sequences which have been determined in large scale sequencing efforts supported by the EC. Gene targeting is even of greater importance for plant breeding since it offers unique opportunities to eliminate undesired traits and to generate transgenic plants with predictable expression levels. Most importantly, this method is not restricted to few model plant species. In contrast to anti-sense approaches it generates predictable and stable phenotypes and gene targeting does not depend on integration preferences of T-DNA or transposable elements.
However, despite of world-wide efforts, no efficient and reproducible protocol for gene targeting could be developed for plants. Probably plant-specific differences in the underlying mechanisms account for this failure and we had not sufficient knowledge on the basic mechanisms and their regulation in plants to change this situation r Recently, however, good candidates of processes (homologous, illegitimate recombination and possibly mismatch repair) were identified which contribute to gene targeting. Therefore we propose to study these processes in plants and manipulate them in a way resulting in an improvement of gene targeting. In detail, we will set up a mutant screen to find mutants affected in the regulation of homologous recombination. Overexpression of recombination-promoting enzymes stimulates homologous recombination and we will exploit this mechanism for improving gene targeting. Yeast will be used as a model system to find genes involved in illegitimate recombination, a process competing with homologous recombination, and the genes will be inactivated in plants.
Strategies for gene targeting will be improved and schemes developed allowing for direct selection of gene replacement. It will be determined whether or not mismatch repair has the same inhibitory effect on homologous recombination and gene targeting as observed in mammalian cells. Gene targeting technology will be applied to inactivate a mismatch repair gene in Arabidopsis and these plants will be analysed. Moreover, the newly developed technology will be transferred from the model plant Arabidopsis to the crop plant Brassica napus and it will be attempted to inactivate a gene of commercial importance.
This proposal is directly relevant for small and medium size enterprises (SME) engaged in plant biotechnology. Its importance is demonstrated by the participation of two SMEs in this project.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences genetics DNA
- agricultural sciences agriculture, forestry, and fisheries agriculture agronomy plant breeding
- natural sciences biological sciences genetics genomes
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Coordinator
CF4 3UH Cardiff
United Kingdom
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