Method of generating .rho.0 cells

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/09 (2006.01)

Patent

CA 2707732

Described is a method of the generation of ~° cells using a mitochondrial targeted restriction endonuclease. This method comprises (a) transfecting cells with an expression vector containing a gene encoding a fusion protein comprising a mitochondrial targeting sequence (MTS) and a restriction endonuclease operatively linked to a suitable promoter; (b) culturing the transfected cells over a sufficient period of time; and (c) selecting ~° cells, e.g., via FACS analysis.

L'invention porte sur un procédé de génération de cellules ?° à l'aide d'une endonucléase de restriction ciblée mitochondriale. Ce procédé comprend les opérations consistant à : (a) transfecter des cellules par un vecteur d'expression contenant un gène codant pour une protéine de fusion comprenant une séquence de ciblage mitochondriale (MTS) et une endonucléase de restriction liée de façon fonctionnelle à un promoteur approprié ; (b) mettre en culture les cellules transfectées sur une période de temps suffisante et (c) sélectionner des cellules ?°, par exemple, par l'intermédiaire d'une analyse par trieur de cellules marquées par fluorescence (FACS).

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