Methods and compositions for generating a genetically...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/00 (2006.01) C12N 15/00 (2006.01)

Patent

CA 2470527

The present invention provides methods and compositions for the integration of a nucleotide sequence of interest into the genome. Specifically, the methods of the present invention comprise contacting a lentiviral vector with the plasma membrane of an oocyte, early stage embryo, or blastula under conditions that permit the entry of the viral vector and the subsequent integration of the nucleotide sequence of interest into the genome. The methods further comprise culturing the oocyte or embryo contacted with the lentiviral vector under conditions that allow for the formation of a pre-implantation embryo. Subsequently, the pre-implantation embryo can be transferred into a recipient vertebrate where it is allowed to develop into at least one genetically modified animal. The methods and compositions of the invention thereby allow for the production of genetically modified animals, particularly, vertebrates, and particularly mammals.

La présente invention concerne des méthodes et des compositions pouvant intégrer une séquence nucléotidique d'intérêt dans le génome. Plus particulièrement, les méthodes de l'invention consistent à placer un vecteur lentiviral au contact de la membrane plasmique d'un ovocyte, d'un embryon au stade précoce ou de blastulas, dans des conditions permettant l'introduction du vecteur viral et l'intégration subséquente de la séquence nucléotidique d'intérêt dans le génome. Ces méthodes consistent en outre à cultiver l'ovocyte ou l'embryon placé au contact du vecteur lentiviral, dans des conditions qui permettent la formation d'un embryon de pré-implantation, lequel est ensuite transféré chez un vertébré receveur où on le laisse se développer en au moins un animal génétiquement modifié. Les méthodes et compositions de l'invention permettent ainsi de produire des animaux génétiquement modifiés, en particulier des vertébrés, et notamment des mammifères.

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