An expression silencing system and different uses thereof

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/82 (2006.01) A01H 5/00 (2006.01) C12N 15/63 (2006.01)

Patent

CA 2359356

The invention relates to an expression-silencing system comprising a first DNA construct comprising a nucleotide sequence corresponding to the T7 RNA polymerase gene (T7-pol) which carries an NLS sequence, and at least one promoter and at least one terminator sequence operably linked to the T7-pol; a second DNA construct comprising a T7 promoter sequence (pT7), at least one targeting sequence downstream to said pT7 and at least one 3' non-translated terminator sequence operably linked to the targeting sequence; which system can, upon its introduction into a cell, substantially silence the expression at the RNA level of a target sequence in the cell, in a tissue or organ regenerated from said cell, or in a progeny thereof, substantially silenced, by causing the substantial disappearance of the RNA or RNA transcript carrying said sequence or a functional part thereof.

L'invention concerne un système de mise au silence de l'expression comprenant un premier produit de recombinaison d'ADN contenant une séquence de nucléotides correspondant au gène d'ARN polymérase T7 (T7-pol) portant une séquence de NLS, au moins une séquence promoteur et au moins une séquence terminale reliées à T7-pol; un deuxième produit de recombinaison d'ADN contenant une séquence promoteur de T7 (pT7), au moins une séquence de ciblage en aval dudit pT7 et au moins une séquence terminale 3' non traduite reliée à la séquence de ciblage. Ce système peut, lorsqu'on l'introduit dans une cellule, mettre pratiquement au silence l'expression au niveau de l'ARN d'une séquence ciblée de la cellule, d'un tissu ou d'un organe régénéré à partir de ladite cellule ou d'une descendance de ladite cellule, étant donné qu'il provoque la disparition de l'ARN ou du transcrit de l'ARN portant ladite séquence ou une de ses parties fonctionnelles.

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