Methods for cloning non-human mammals using reprogrammed donor chromatin or donor cells
First Claim
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1. A method of cloning a non-human mammal, said method comprising the steps of:
- (a) permeabilizing a non-human mammalian cell, thereby generating a permeabilized cell having pores in its plasma membrane or a partial plasma membrane;
(b) incubating said permeabilized cell in an extract from a mitotic cell under conditions that allow chromatin condensation and nuclear envelope breakdown of said permeabilized cell;
(c) inserting said cell formed in step (b) into a nucleated or enucleated oocyte, thereby forming a reconstituted oocyte; and
(d) transferring said reconstituted oocyte or an embryo formed from said reconstituted oocyte into the uterus of a host mammal under conditions that allows said reconstituted oocyte or said embryo to develop into a fetus.
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Abstract
The invention provides methods for cloning mammals that allow the donor chromosomes or donor cells to be reprogrammed prior to insertion into an enucleated oocyte. The invention also features methods of inserting chromosomes or nuclei into recipient cells.
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Citations
24 Claims
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1. A method of cloning a non-human mammal, said method comprising the steps of:
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(a) permeabilizing a non-human mammalian cell, thereby generating a permeabilized cell having pores in its plasma membrane or a partial plasma membrane; (b) incubating said permeabilized cell in an extract from a mitotic cell under conditions that allow chromatin condensation and nuclear envelope breakdown of said permeabilized cell; (c) inserting said cell formed in step (b) into a nucleated or enucleated oocyte, thereby forming a reconstituted oocyte; and (d) transferring said reconstituted oocyte or an embryo formed from said reconstituted oocyte into the uterus of a host mammal under conditions that allows said reconstituted oocyte or said embryo to develop into a fetus. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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Specification