Methods of modifying eukaryotic cells

Details Methods of modifying eukaryotic cells Methods of modifying eukaryotic cells

C - Chemistry, Metallurgy

12

N

C12N 15/85 (2006.01) A01K 67/027 (2006.01) C12N 15/90 (2006.01)

Patent

CA 2426398

A method for engineering and utilizing large DNA vectors to target, via homologous recombination, and modify, in any desirable fashion, endogenous genes and chromosomal loci in eukaryotic cells. These large DNA targeting vectors for eukaryotic cells, termed LTVECs, are derived from fragments of cloned genomic DNA larger than those typically used by other approaches intended to perform homologous targeting in eukaryotic cells. Also provided is a rapid and convenient method of detecting eukaryotic cells in which the LTVEC has correctly targeted and modified the desired endogenous genes(s) or chromosomal locus (loci) as well as the use of these cells to generate organisms bearing the genetic modification.

L'invention concerne un procédé de manipulation et d'utilisation de grands vecteurs d'ADN afin de cibler, via une recombinaison homologue, et modifier, d'une façon quelconque souhaitée, des gène endogènes et des loci chromosomiques dans des cellules eucaryotes. Ces grands vecteurs d'ADN de ciblage pour cellules eucaryotes, dénommés LTVEC, sont dérivés de fragments d'ADN génomique cloné plus grands que ceux habituellement utilisés dans d'autres approches cherchant à réaliser un ciblage homologue dans des cellules eucaryotes. L'invention concerne aussi un procédé rapide et adéquat de détection de cellules eucaryotes dans lesquelles le LTVEC a correctement ciblé et modifié le gène endogène ou le locus chromosomique recherché, ainsi que l'utilisation de ces cellules afin de produire des organismes portant la modification génétique.

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