Methods and compositions for transforming cells

C - Chemistry – Metallurgy – 12 – N

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C12N 15/90 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C07K 14/805 (2006.01) C12N 15/64 (2006.01) C12N 15/86 (2006.01) C12N 15/867 (2006.01)

Patent

CA 2258007

Methods and compositions for transforming cells, resulting in efficient and stable site-specific integration of transgenes, are disclosed. Transformation is achieved by introducing into a cell an acceptor vector, preferably a retroviral vector, which integrates into the genome of the cell. The acceptor vector comprises two incompatible lox sequences, L1 and L2. A donor vector is then introduced into the cell comprising a transgene flanked by the same L1 and L2 sequences. Stable gene transfer is initiated by contacting the lox L1 and L2 sequences with Cre recombinase.

Procédés et compositions permettant de transformer des cellules, qui entraînent une intégration spécifique au site, efficace et stable de transgènes. La transformation s'obtient par l'introduction dans une cellule d'un vecteur accepteur, de préférence un vecteur rétroviral, qui s'intègre dans le génome de la cellule. Ledit vecteur accepteur comporte deux séquences lox incompatibles, L1 et L2. Un vecteur donneur est ensuite introduit dans la cellule comportant le transgène flanqué des mêmes séquences L1 et L2. On parvient à un transfert de gène stable en mettant en contact les séquences lox L1 et L2 avec de la recombinase Cre.

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