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US 6,376,743 B1Granted
Mammalian transgenesis by intracytoplasmic sperm injection
AssigneeUniversity of Hawaii
InventorRyuzo Yanagimachi
Life of this patent
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Abstract
Coinjection of unfertilized mouse oocytes with sperm heads and exogenous nucleic acid encoding a transgene results in transgene-expressing embryos, reflecting nucleic acid-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos resulting from coinjection produced offspring expressing the integrated transgene.
Claims20
A method for obtaining a transgenic embryo, comprising the steps of:
- incubating a nucleic acid that is exogenous to the embryo with a membrane-disrupted sperm head or a demembranated sperm head for a period of time;
- co-inserting the exogenous nucleic acid and sperm head into an unfertilized oocyte to form a transgenic fertilized oocyte;
- and
- allowing the transgenic fertilized oocyte to develop into a transgenic embryo.
1 more element
A method for obtaining a transgenic embryo, comprising the steps of:
- obtaining a membrane-disrupted sperm head or a demembranated sperm head;
- mixing the membrane-disrupted sperm head or demembranated sperm head with a nucleic acid that is exogenous to the embryo;
- co-inserting the mixture into an isolated unfertilized metaphase II oocyte to form a transgenic fertilized oocyte;
- and
- allowing the transgenic fertilized oocyte to develop into a transgenic embryo.
2 more elements
Dependencies are inferred from the claim text.