Dna construct for effecting homologous recombination and...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/67 (2006.01) C07K 14/505 (2006.01) C07K 19/00 (2006.01) C12N 15/18 (2006.01) C12N 15/62 (2006.01) C12N 15/63 (2006.01) C12N 15/85 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2190289

The invention relates to constructs comprising: a) a targeting sequence; b) a regulatory sequence; c) an exon; and d) an unpaired splice-donor site. The invention further relates to a method of producing protein in vitro or in vivo comprising the homologous recombination of a construct as described above within a cell. The homologously recombinant cell is then maintained under conditions which will permit transcription and translation, resulting in protein expression. The present invention further relates to homologously recombinant cells, including primary, secondary, or immortalized vertebrate cells, methods of making the cells, methods of homologous recombination to produce fusion genes, methods of altering gene expression in the cells, and methods of making a protein in a cell employing the constructs of the invention.

Produits de recombinaison comprenant: a) une séquence de ciblage; b) une séquence régulatrice; c) un exon; d) un site donneur-épissage non apparié. L'invention se rapporte également à un procédé de production de protéines in vitro ou in vivo, comprenant la recombinaison homologue à l'intérieur d'une cellule d'un produit de recombinaison tel que décrit ci-dessus. La cellule de recombinaison homologue est maintenue par la suite dans des conditions permettant la transcription et la traduction, afin d'obtenir l'expression de la protéine. L'invention se rapporte en outre à des cellules de recombinaison homologue, comprenant des cellules de vertébré primaires, secondaires ou immortalisées, à des procédés de production de ces cellules, à des procédés de recombinaison homologue permettant de produire des gènes de fusion, à des procédés de modification de l'expression génique dans les cellules, ainsi qu'à des procédés permettant de produire une protéine dans une cellule utilisant les produits de recombinaison décrits.

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