Method for integrating genes at specific sites in mammalian...

Details Method for integrating genes at specific sites in mammalian... Method for integrating genes at specific sites in mammalian...

C - Chemistry, Metallurgy

12

N

C12N 15/90 (2006.01) C07K 14/705 (2006.01) C07K 16/28 (2006.01) C12N 5/10 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2283740

A method for achieving site specific integration of a desired DNA at a target site in a mammalian cell via homologous recombination is described. This method provides for the reproducible selection of cell lines wherein a desired DNA is integrated at a predetermined transcriptionally active site previously marked with a marker plasmid. The method is particularly suitable for the production of mammalian cell lines which secrete mammalian proteins at high levels, in particular immunoglobulins. Vectors and vector combinations for use in the subject cloning method are also provided.

Procédé pour assurer l'intégration spécifique à un site d'un ADN voulu dans un site cible d'une cellule de mammifère par recombinaison homologue. Ce procédé qui permet une sélection reproductible de lignées cellulaires, consiste en l'intégration d'un ADN voulu à un site prédéterminé actif du point de vue transcriptionnel, marqué préalablement au moyen d'un plasmide de marquage. Ce procédé est particulièrement adapté à la production de lignées cellulaires de mammifères, sécrétant des quantités importantes de protéines de mammifères, en particulier des immunoglobulines. L'invention porte en outre sur des vecteurs et des combinaisons de vecteurs utilisables dans le procédé de clonage de l'invention.

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