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US 7,294,754 B2Granted
Method for generating an animal homozygous for a genetic modification
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsWilliam Poueymirou, Thomas M. Dechiara, Wojtek Auerbach, David Frendewey, David M. Valenzuela
Life of this patent
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Abstract
Methods of generating modified embryos and mammals by introduction of donor cells into an early stage embryo are provided, such that the resulting embryo and animal generated therefrom has a significant or complete contribution to all tissues from the donor cells and is capable of transmitting the donor cell DNA.
Claims15
A method for generating a mouse homozygous for a genetic modification, comprising:
- (a) introducing a female mouse embryonic stem (ES) donor cell into a pre-morula diploid mouse host embryo, wherein the ES donor cell is heterozygous for the genetic modification and is obtained from an inbred strain;
- (b) culturing the embryo of (a) to the blastocyst stage;
- (c) introducing the embryo of (b) into a surrogate mother for gestation, wherein a modified female mouse is generated having at least 90% cellular contribution from the female donor cell;
- (a′) introducing a male mouse ES donor cell into a pre-morula diploid mouse host embryo, wherein the ES donor cell is heterozygous for the genetic modification and is obtained from an inbred strain;
- (b′) culturing the embryo of (a′) to the blastocyst stage;
- (c′) introducing the embryo of (b′) into a surrogate mother for gestation, wherein a modified male mouse is generated;
- and
- (d) breeding a sexually mature modified male mouse and a sexually mature modified female mouse of (c and c′), wherein a mouse homozygous for the genetic modification is generated.
5 more elements
Dependencies are inferred from the claim text.