Use of anti-sense sequences to increase responsiveness to...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/67 (2006.01) C07K 14/54 (2006.01) C07K 14/555 (2006.01) C07K 14/705 (2006.01) C07K 14/715 (2006.01) C12N 5/10 (2006.01) C12N 15/11 (2006.01) C12N 15/12 (2006.01) C12N 15/85 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2235962

A method for regulating the production level of a desired protein in a transformed mammalian cell, comprising: a directly amplifiable gene sequence, in expressible form, which when expressed renders said cell more resistant to a toxic agent; a gene of interest, in expressible form, which encodes a desired protein foreign to said cell; and an anti-sense gene sequence, in transcribable form, encoding an anti-sense RNA capable of selectively hybridizing to at least a portion of the mRNA transcribed from said directly amplifiable gene sequence so as to inhibit the translation of said mRNA, such that a higher level of amplification of said directly amplifiable gene and said gene of interest can be achieved, if the cell is exposed to a sufficient level of the toxic agent, than in the absence of said anti-sense RNA.

L'invention concerne un procédé pour réguler le niveau de production d'une protéine souhaitée dans une cellule mammalienne transformée, faisant appel à: une séquence d'un gène directement amplifiable, sous une forme permettant son expression, qui, lors de l'expression, rend la cellule plus résistante à un agent toxique; un gène utile, sous une forme permettant son expression, codant pour une protéine souhaitée étrangère à la cellule; et une séquence d'un gène anti-sens, sous une forme se prêtant à une transcription, codant pour un ARN anti-sens capable de former sélectivement un hybride avec au moins une portion de l'ARNm obtenu par la transcription de ladite séquence de gène directement amplifiable, ce qui inhibe la traduction dudit ARNm, de sorte qu'un niveau d'amplification plus élevé dudit gène directement amplifiable et dudit gène utile peut être obtenu, lorque la cellule est exposée à un niveau suffisant de l'agent toxique, qu'en l'absence dudit ARN anti-sens.

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