Novel modified nucleic acid sequences and methods for...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/30 (2006.01) A01K 67/027 (2006.01) A61K 48/00 (2006.01) C07K 14/445 (2006.01) C07K 14/47 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/67 (2006.01) C12N 15/85 (2006.01) A61K 39/00 (2006.01)

Patent

CA 2306799

The invention provides modified recombinant nucleic acid sequences (preferably DNA) and methods for increasing the mRNA levels and protein expression of proteins which are known to be, or are likely to be, difficult to express in cell culture systems, mammalian cell culture systems, or in transgenic animals. The preferred "difficult" protein candidates for expression using the recombinant techniques of the invention are those proteins derived from heterologous cells preferably those of lower organisms such as parasites, bacteria, and virus, having DNA coding sequences comprising high overall AT content or AT rich regions and/or mRNA instability motifs and/or rare codons relative to the recombinant expression system to be used.

L'invention concerne des séquences modifiées d'acides nucléiques de recombinaison (ADN, de préférence) et des procédés permettant d'accroître les niveaux d'ARNm et l'expression des protéines, dont l'expression est notoirement ou vraisemblablement difficile dans les systèmes de culture de cellules, les systèmes de culture de cellules mammaliennes, ou chez les animaux transgéniques. Les protéines "difficiles" préférées qui sont candidates à l'expression, via les techniques de recombinaison décrites dans l'invention, sont les protéines dérivées de cellules hétérologues. De préférence, il s'agit des cellules d'organismes inférieurs comme les parasites, les bactéries et les virus ayant des séquences de codage ADN à teneur globalement élevée en adénine/thymine (AT) ou comportant des zones riches en AT et/ou des motifs d'instabilité d'ARNm et /ou des codons rares par rapport au système d'expression de recombinaison à utiliser.

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