A novel method for the integration of foreign dna into...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/52 (2006.01) A01H 1/00 (2006.01) A01H 5/00 (2006.01) C07K 19/00 (2006.01) C12N 5/10 (2006.01) C12N 9/00 (2006.01) C12N 15/62 (2006.01) C12N 15/82 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2306188

Compositions and methods for introducing a DNA of interest into a genomic target site are provided. In particular, the methods and compositions involve the use of a combination of target sites for two site specific recombinases and expression of a chimeric recobminase with dual target site specificity. Thus, the compositions comprise novel site-specific recombinases with specificities to multiple target sites, and nucleotide sequences and expression cassettes encoding these recombinases or target sites. The methods involve transforming a eukaryotic cell having target sites for the novel recombinase with a DNA of interest that is flanked by corresponding target sites. Expression of the recombinase results in integration of the DNA of interest into the genome of the cell. The compositions and methods of the invention have use in the construction of stably transformed eukaryotic cells, and in particular, plant cells. The methods result in the efficient targeted genomic intergration of DNA by site-specific recombination.

Cette invention concerne des compositions et des procédés permettant d'introduire un AND spécifique dans un site cible génomique; les procédés et les compositions de cette invention impliquant l'utilisation d'une combinaison de sites cibles pour deux recombinases spécifiques et l'expression d'une recombinase chimère ayant une spécificité à double site cible. Les compositions contiennent par conséquent de nouvelles recombinases à spécificité de site ayant des spécificités pour des sites cibles multiples, ainsi que des séquences nucléotidiques et des cassettes d'expression codant ces recombinases ou ces sites cibles. Les procédés impliquent la transformation d'une cellule eucaryote comportant des sites cibles pour la nouvelle recombinase avec un AND spécifique qui est entouré de sites cibles correspondants. L'expression de la recombinase produit l'intégration de l'ADN spécifique dans le génome de la cellule. Les compositions et les procédés de cette invention sont utiles dans la construction de cellules eucaryotes transformées de manière stable et plus particulièrement dans la construction de cellules de plantes. Les procédés présentés permettent d'effectuer l'intégration génomique ciblée et efficace d'ADN par recombinaison dirigée.

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