Methods for creating transgenic animals
AssigneeWisconsin Alumni Research Foundation
InventorsRobert D. Bremel, Anthony W. S. Chan, Jane C. Burns
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Abstract
The present invention provides improved methods and compositions for the generation of transgenic non-human animals. The present invention permits the introduction of exogenous nucleic acid sequences into the genome of unfertilized eggs (e.g., pre-maturation oocytes and pre-fertilization oocytes) by microinjection of infectious retrovirus into the perivitelline space of the egg. The methods of the present invention provide an increased efficiency of production of transgenic animals with a reduced rate of generating animals which are mosaic for the presence of the transgene.
Claims16
A method for introducing a polynucleotide contained within the genome of a recombinant retrovirus into the genome of a bovine unfertilized oocyte, comprising:
- a) providing:
- i) a bovine unfertilized egg comprising an oocyte having a plasma membrane and a zona pellucida, said plasma membrane and said zona pellucida defining a perivitelline space;
- ii) an aqueous solution comprising a polynucleotide contained within the genome of a recombinant retrovirus;
- and
- b) introducing said solution comprising said recombinant retrovirus into said perivitelline space under conditions which permit the integration of said polynucleotide contained within the genome of the recombinant retrovirus into the genome of said oocyte.
A method for the production of a transgenic bovine embryo comprising:
- a) providing:
- i) an unfertilized bovine egg comprising an oocyte having a plasma membrane and a zona pellucida, said plasma membrane and said zona pellucida defining a perivitelline space;
- ii) an aqueous solution containing infectious retrovirus carrying a gene encoding a protein of interest;
- b) introducing said solution containing infectious retrovirus into said perivitelline space under conditions which permit the infection of said oocyte, thereby giving an infected oocyte;
- c) contacting said infected oocyte with sperm under conditions which permit the fertilization of said infected oocyte to produce a bovine embryo, and
- d) verifying the presence of the gene from the infectious retrovirus in the embryo, the bovine animal or its offspring.
Dependencies are inferred from the claim text.