Eukaryotic cells stably expressing genes from multiple...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/10 (2006.01) C12N 15/10 (2006.01) C12N 15/64 (2006.01)

Patent

CA 2324518

A method is described for producing recombinant eukaryotic cell lines expressing multiple proteins or RNAs of interest. Eukaryotic host cells are transfected with (a) a first episome which contains a sequence that promotes autonomous replication of the episome in the cells and a first gene encoding a protein of interest; and (b) a second episome containing a sequence that promotes autonomous replication of the episome in the cells and a second gene encoding a protein of interest. Transfected cells are obtained expressing one or more proteins that promote nuclear retention of the episomes. The cells are grown under conditions wherein the episomes express the first and second genes.

L'invention concerne un procédé de production de lignées cellulaires eucaryotes recombinées exprimant de multiples protéines ou ARN d'intérêt. On transfecte des cellules hôtes eucaryotes avec (a) un premier épisome contenant une séquence favorisant la réplication autonome de l'épisome dans ces cellules et un premier gène codant une protéine d'intérêt; et (b) un second épisome contenant une séquence favorisant la réplication autonome de l'épisome dans ces cellules et un second gène codant une protéine d'intérêt. On obtient des cellules transfectées en exprimant une ou plusieurs protéines favorisant la rétention nucléaire des épisomes. On développe ces cellules dans des conditions où les épisomes expriment les premier et second gènes.

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