Non-bacterial cloning in delivery and expression of nucleic...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) C12N 5/10 (2006.01) C12N 7/01 (2006.01) C12N 9/00 (2006.01) C12N 15/11 (2006.01) C12N 15/85 (2006.01) C12N 15/86 (2006.01)

Patent

CA 2315242

Novel double-stranded DNAs, expression vectors and methods for their use are provided in which the intracellular expression of the double-stranded DNAs alters the phenotype of a cell to determine the function of a gene of interest. The double-stranded DNAs encode a family of catalytic RNAs targeted to the mRNA of a gone of interest. Cleavage of the mRNA results in an altered cell phenotype from which the function of the product encoded by the mRNA is determined. The compositions find use in high-throughput screens to assign gene functions which eliminate the requirement of bacterial cloning.

L'invention concerne de nouveaux ADN double brin, des vecteurs d'expression, et leurs méthodes d'utilisation, l'expression intracellulaire de ces ADN double brin modifiant le phénotype d'une cellule de manière à déterminer la fonction d'un gène d'intérêt. Ces ADN double brin codent une famille d'ARN catalytiques ciblés vers l'ARN messager d'un gène d'intérêt. On obtient, par clivage de l'ARN messager, un phénotype cellulaire modifié, à partir duquel on peut déterminer la fonction du produit codé par cet ARN messager. Les compositions de la présente invention sont notamment utiles dans des cribles à haut rendement, permettant d'attribuer des fonctions géniques ne nécessitant aucun clonage bactérien.

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