Genetic complementation compositions and methods
AssigneeThe Regents of the University of California
InventorsMaci L. MUELLER, Alba Veronica LEDESMA, Pablo Juan ROSS, Alison Louise Van EENENNAAM
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Abstract
Described in several exemplary embodiments herein are germline complementation methods and compositions, particularly NANOS3 deficient cells and non-human animals. In some embodiments, the NANOS3 deficient non-human animals are germline ablated.
Claims39
A complemented non-human animal or embryo comprising:
- a first population of cells comprising one or more cells, wherein the first population of cells consists of an engineered non-human animal cell or population thereof comprising a NANOS3 gene modification, wherein the NANOS3 gene modification reduces or eliminates the expression of a NANOS3 gene product;
- a second population of cells comprising one or more cells, wherein the second population of cells are not an engineered non-human cell or population thereof comprising a comprising a NANOS3 gene modification, wherein the NANOS3 gene modification reduces or eliminates the expression of a NANOS3 gene product.
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An engineered non-human animal cell or population thereof comprising:
- a NANOS3 gene modification, wherein the NANOS3 gene modification reduces or eliminates the expression of a NANOS3 gene product, wherein
- the NANOS3 gene modification is
- (i) an insertion of one or more nucleotides;
- (ii) a deletion of one or more nucleotides;
- (iii) a substitution of one or more nucleotides;
- or
- (iv) any combination of (i)-(iii).
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A method of generating a NANOS3 modified non-human animals or embryos, the method comprising:
- introducing one or more NANOS3 gene modifications to a non-human animal cell, wherein the NANOS3 gene modification reduces or eliminates the expression of a NANOS3gene product;
- and
- one or more of the following techniques:
- somatic cell nuclear transfer, oocyte pronuclear DNA microinjection, zygote microinjection, or embryo microinjection, intracytoplasmic sperm injection, in vitro fertilization, embryo transfer, in vitro embryo culture, or any combination thereof.
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A method of non-human animal embryo complementation comprising:
- introducing a self-renewing exogenous population of cells into a non-human animal preimplantation embryo, optionally at about day 3, 4, 5, or 6 post fertilization;
- optionally washing the non-human animal preimplantation embryo in HEPES or other suitable buffe;
- and
- culturing the non-human preimplantation embryo in a suitable culture media optionally consisting of a 1:1 ratio by volume of a suitable bovine culture media that is at least supplemented with N2, B27, FGF, and IWR-1.
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Dependencies are inferred from the claim text.