Method of performing transgenesis

Details Method of performing transgenesis Method of performing transgenesis

A - Human Necessities

01

K

A01K 67/027 (2006.01) A01K 67/033 (2006.01) C12N 15/00 (2006.01) C12N 15/87 (2006.01) C12N 15/89 (2006.01)

Patent

CA 2340242

The invention provides a method for generating transgenic animals and cells by the coinsertion of nucleic acid and a nucleus into an unfertilized oocyte. Preferably, the coinsertion is by microinjection and more preferably by piezo- electrically actuated microinjection. Transgene (tg) expressing embryos are here produced following coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or .beta.-galactosidase reporter. The microinjected oocyte may be allowed to develop into differentiated cells or stem cells; into an embryo in vitro prior to transfer into a host surrogate mother; or it may be transferred directly into a host surrogate mother. Embryonic development can occur to term, such that the offspring possess transgenic modifications that may alter their characteristics (phenotype) and are, in turn, transmitted to their offspring.

L'invention concerne un procédé de production d'animaux et de cellules transgéniques par la co-insertion d'acide nucléique et d'un noyau dans un ovocyte non fécondé. De préférence, la co-insertion est effectuée par micro-injection, et idéalement par micro-injection piézo-électrique. Des embryons exprimant le transgène (tg) sont produits après co-injection d'ovocytes de souris non fécondés à l'aide de têtes de sperme et d'ADN exogène codant pour une protéine fluorescente verte (GFP) ou pour un rapporteur de galactosidase .beta.. On peut laisser se développer l'ovocyte micro-injecté en cellules différenciées ou en cellules souches ; en embryon in vitro avant transfert dans une mère porteuse hôte ; ou on peut le transférer directement dans une mère porteuse hôte. Le développement embryonnaire peut se faire à terme, de sorte que la progéniture acquiert des modifications transgéniques pouvant modifier ses caractéristiques (phénotype), et qui peuvent ensuite être transmises à la descendance.

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