Novel sirna libraries and their production and use

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/74 (2006.01) C12N 15/10 (2006.01) C12N 15/11 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2493564

The present invention relates to methods and compositions for the elucidation of gene function and the identification of novel genes. Specifically, the present invention relates to methods and compositions for improved functional genomic screening, functional inactivation of specific essential or non- essential genes, and identification of genes that are modulated in response to specific stimuli or encode recognizable phenotypic traits. In particular, the compositions of the present invention include, but are not limited to, expression cassettes comprising a novel dual promoter transcription system, that utilizes modified promoters, preferably containing complementary termination sequences, positioned across a coding sequence and in opposite orientation to each other. In addition, the present invention includes libraries comprising the expression cassettes of the invention, including vectors for transforming cells, such as replication-deficient retroviral vectors. The invention also includes methods for the production and screening of dsRNA/siRNA libraries, as well as therapeutic uses for the siRNAs expressed in accordance with the invention.

La présente invention se rapporte à des procédés et à des compositions permettant d'élucider la fonction de gênes et d'identifier de nouveaux gênes. Spécifiquement, la présente invention se rapporte à des procédés et à des compositions permettant d'effectuer un criblage génomique fonctionnel amélioré, une inactivation fonctionnelle de gênes essentiels ou non essentiels spécifiques, ainsi que l'identification de gênes qui sont modulés en réponse à des stimuli spécifiques ou qui codent des traits phénotypiques reconnaissables. En particulier, les compostions de la présente invention incluent, non exclusivement, des cassettes d'expression comportant un nouveau système de transcription promoteur double, qui utilise des promoteurs modifiés, de préférence contenant des séquences de terminaison complémentaires, positionnés à travers une séquence de codage et orientés de manière opposée les uns par rapport aux autres. En outre, la présente invention se rapporte à des banques comportant les cassettes d'expression ci-dessus mentionnées, notamment des vecteurs de transformation de cellules, tels que des vecteurs rétroviraux déficients pour la réplication. L'invention se rapporte également à des procédés de production et de criblage de banques génomiques d'ARN double brin (dsRNA)/de petits fragments d'ARN interférents (siRNA), ainsi qu'à des utilisations thérapeutiques de ces petits fragments d'ARN interférents exprimés conformément à l'invention.

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