Method of nuclear transfer
OwnerIan Lewis, Gabor Vajta +1inventors
InventorsIan Lewis, Gabor Vajta, Tayfur Tecirlioglu
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Abstract
The present invention relates to nuclear transfer methods and embryos developed therefrom. In particular, the present invention relates to a method of nuclear transfer comprising the step of transferring a somatic cell or somatic cell nuclei into a zona pellucida-free, enucleated oocyte.
Claims61
A method of nuclear transfer comprising the step of transferring of a somatic cell or somatic cell nuclei into a zona pellucida-free, enucleated oocyte.
A method for producing a genetically engineered or transgenic non-human mammal comprising the step of:
- (i) inserting, removing or modifying a desired gene or genes in a non-human somatic cell or cell nucleus;
- (ii) inserting the non-human somatic cell or cell nucleus into a zona pellucida-free, enucleated oocyte under conditions suitable for the formation of a reconstituted cell;
- (iii) activating the reconstituted cell to form an embryo;
- (iv) culturing said embryo until greater than the 2-cell developmental stage;
- and
- (v) transferring said cultured embryo to a host mammal such that the embryo develops into a transgenic fetus.
A method for cloning a non-human mammal comprising the steps of:
- (i) inserting a desired non-human somatic cell or cell nucleus into a zona pellucida-free, enucleated oocyte, under conditions suitable for the formation of a reconstituted cell;
- (ii) activating the reconstituted cell to form an embryo;
- (iii) culturing said embryo until greater than the 2-cell developmental stage;
- and
- (iv) transferring said cultured embryo to a host mammal such that the embryo develops into a fetus.
A method according to 31, further comprising the step of attaching the zona pellucida-free, enucleated oocyte to the non-human somatic cell or cell nucleus before fusion.
A method according to any one of claims 2 to 48, further comprising the step of fusing one or more further zona pellucida-free oocytes to the reconstituted cell.
A method for cloning a non-human mammal comprising the steps of:
- (i) inserting a desired non-human somatic cell or cell nucleus into a first zona pellucida-free, enucleated mammalian oocyte, under conditions suitable for the formation of a reconstituted cell;
- (ii) fusing a second or more oocyte to said reconstituted cell thereby increasing the cytoplasmic volume;
- (iii) activating the reconstituted cell to form an embryo;
- (iv) culturing said embryo until greater than the 2-cell developmental stage;
- and
- (v) transferring said cultured embryo to a host mammal such that the embryo develops into a fetus.
A method of culturing a reconstituted cell (embryo) comprising the steps of:
- (i) inserting a desired non-human somatic cell or cell nucleus into a zona pellucida-free, enucleated oocyte, under conditions suitable for the formation of a reconstituted cell (embryo);
- (ii) activating the reconstituted cell;
- (iii) incubating and culturing one or more of said cells until embryos of between 8 to 128 cells develop.
Dependencies are inferred from the claim text.