Stable and long-lasting sirna expression vectors and the...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/869 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2596832

The invention relates to a siRNA expression vector that can inhibit or eliminate the expression of a target gene in a mammalian cell, said vector comprising: a bacterial cassette containing a bacterial origin of replication and a bacterial selection marker M1; a eucaryotic cell selection cassette comprising a marker M2 for selecting eucaryotic cells under the control of an appropriate promoter; a cassette EBV comprising at least one fragment of the antigen EBNA-1, at least one fragment FR, and at least one fragment of the region DYAD; and a siRNA transcription cassette comprising at least one region coding for a siRNA corresponding to the target gene to be inhibited or eliminated, under the control of elements for regulating transcription in mammalian cells, said regulating elements including at least one promoter capable of transcribing a siRNA in mammalian cells, and a transcription terminator. The invention also relates to the applications of one such expression vector.

Vecteur d'expression d'un siRNA, capable d'inhiber ou d'éteindre l'expression d'un gène cible dans une cellule de mammifère, qui comprend : une cassette bactérienne comprenant une origine de réplication bactérienne et un marqueur de sélection bactérienne M1, une cassette de sélection dans les cellules eucaryotes comprenant un marqueur de sélection M2 de cellules eucaryotes sous le contrôle d'un promoteur approprié, une cassette EBV comprenant au moins un fragment de l'antigène EBNA-1, au moins un fragment FR et au moins un fragment de la région DYAD et une cassette de transcription d'un siRNA comprenant au moins une région codant pour un siRNA correspondant au gène cible à inhiber ou à éteindre, sous le contrôle d'éléments régulateurs de transcription dans des cellules de mammifère, lesquels éléments régulateurs incluent au moins un promoteur capable de transcrire un siRNA dans des cellules de mammifère et un terminateur de transcription, et ses applications.

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