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Development of highly specific enzymes for genome manipulation

Ziel

Efficient exploitation of genome information will, among others, depend on our ability, to sequence-specifically modify genomic DNA in vivo. A consortium of 8 academic and 3 industrial units will engineer end nucleases and DNA methyltransferases of programmable specificities which can cleave or methyl ate a single site in a DNA as complex as the human genome. The ability of the enzymes to induce gene replacement or gene silencing by DNA cleavage or cytosine-methylamine, respectively, at predetermined sites will be explored. The project involves coupling of oligonucleotides (Dons) and peptide nucleic acids (Pans) to the N-or C-termini of the C5-methyltransterase Messy and a single-chain derivative of the restriction end nuclease Purim. The covalently attached Dons and Pans will serve as affinity tags to direct the enzyme to specific sequences in the genome. Methods will be developed to introduce the enzymes into cells. Conditions for avoiding enzyme action at non-specific sites will be explored. Development of enzymes of such high specificity should greatly enhance our ability to manipulate genomes and may lead to medical applications e.g. in gene therapy.

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Koordinator

FOUNDATION OF RESEARCH AND TECHNOLOGY - HELLAS
EU-Beitrag
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Adresse
Vassilika Vouton
71110 IRAKLION
Griechenland

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Beteiligte (10)