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US 2026/0193677 A1Published application
Methods of genome editing oocytes
AssigneeThe Regents of the University of California
InventorsAlison Louise Van EENENNAAM, Jason LIN
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Abstract
Provided in this disclosure are methods of producing a fertilized embryo comprising a genomic edit and engineered cells and organisms therefrom. In certain aspects, engineered cells can be created utilizing AAVs and CRISPR/Cas systems.
Claims21
A method of producing a fertilized embryo comprising a genomic edit, the method comprising:
- a) providing isolated mature cumulus-oocyte complexes (COC) from a first non-human organism;
- b) removing the cumulus cells from the isolated mature COC to produce denuded oocytes;
- c) incubating the denuded oocytes with sperm, adeno-associated virus (AAV) particles comprising a donor nucleic acid having homology to a nucleotide sequence adjacent to a target genomic site, and supplemental mature COC from a second non-human organism to produce a first plurality of cells comprising at least one zygote comprising the donor nucleic acid;
- d) removing the cumulus cells from the first plurality of cells thereby producing a plurality of denuded cells comprising at least one denuded zygote;
- e) electroporating the plurality of denuded cells in the presence of a site-directed nuclease that binds to the target genomic site;
- and
- f) culturing the electroporated plurality of denuded cells to produce at least one fertilized embryo comprising the genomic edit.
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Dependencies are inferred from the claim text.