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Targeted gene integration in plants: vectors, mechanisms and applications for protein production


Plants can be considered as natural, solar energy powered, and environmentally friendly factories that can be improved for production of proteins with therapeutical value. However, current transgenic technologies are not accurate enough to secure reliable and precise protein production. Random chromosomal integration of transgenic inserts often causes misregulation of their transcription including complete gene silencing. Therefore the ability to modify plant genomes in a precise manner is arguably the most important missing technology in the toolkit of plant biologists. DNA integration, via homologous recombination (Gene targeting - GT) has proved a powerful technology in many species. It enables disruption, modification or replacement of an endogenous locu s by a new gene, in the native genomic context. Despite its success in other organisms, GT has not yet become a routine technique in plants. New findings, several of which from the TAGIP partners, suggest that GT became a realistic goal. To achieve this go al we propose to gain fundamental knowledge in the field of genome maintenance/modifications via DNA recombination and to apply this knowledge to develop new technologies for precise engineering of plant genomes. Finally, this knowledge will be harnessed f or the production of proteins of high value. Work will be carried in Arabidopsis and in model crop plants. The objectives of TAGIP are: -Development of new vectors and delivery methods for GT -Altering DNA recombination pathways to enhance GT -Application of GT for production of therapeutic proteins in plants The GT mechanism is a fundamental and open question. The tools and knowledge to be developed in TAGIP are relevant to agriculture (two small Agbiotech companies are active participants in the consortiu m) and medicine. The project is of particular importance for the EU states where public concern over the use of GMOs has generated a strong need for controlled integration of beneficial transgenic

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