C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 5/10 (2006.01) A01K 67/027 (2006.01) A61D 19/04 (2006.01) A61K 35/54 (2006.01) C12N 5/16 (2006.01) C12N 15/06 (2006.01) C12N 15/87 (2006.01) C12N 15/89 (2006.01)
Patent
CA 2229657
A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.
Ce procédé de reconstitution d'un embryon animal consiste à transférer un noyau diploïde dans un ovocyte où l'on a stoppé la métaphase de la seconde division méiotique. Cet ovocyte n'est pas activé au moment du transfert de manière à ce que le noyau donneur puisse être exposé, pendant une certaine période, au cytoplasme receveur. Le noyau diploïde peut être donné par une cellule dans l'une ou l'autre des phases G0 ou G1 du cycle cellulaire, au moment du transfert. Ensuite, on active l'embryon reconstitué. On maintient une ploïdie correcte pendant l'activation, par exemple, en incubant l'embryon reconstitué en présence d'un inhibiteur de microtubules tel que le nocodazole. L'embryon reconstitué peut donner naissance à un ou plusieurs animaux vivants. L'invention est utile dans la production d'animaux transgéniques de même que dans celle d'animaux non transgéniques présentant un grand intérêt génétique.
Campbell Keith Henry Stockman
Wilmut Ian
Roslin Institute (edinburgh)
Sim & Mcburney
LandOfFree
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