Method for making genetic modifications
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 contribution to all tissues from the donor cells and is capable of transmitting the donor cell DNA.
Claims18
A method for making a mouse embryo homozygous for a genetic modification, comprising:
- (a) incorporating a female mouse ES donor cell into a first pre-morula diploid mouse host embryo, wherein the female mouse ES donor cell is heterozygous for the genetic modification and is from an inbred strain;
- (b) culturing the first embryo to the blastocyst stage;
- (c) gestating the embryo of (b) in a surrogate mother, wherein upon gestation a modified female mouse is obtained having at least 90% cellular contribution from the female mouse ES donor cell;
- (a′) incorporating a male mouse ES donor cell into a second pre-morula diploid mouse host embryo, wherein the male mouse ES donor cell is heterozygous for the genetic modification and is from an inbred strain;
- (b′) culturing the embryo of (a′) to the blastocyst stage;
- (c′) gestating the embryo of (b′) in a surrogate mother, wherein upon gestation a modified male mouse is obtained having at least 90% cellular contribution from the male mouse ES donor cell;
- and,
- (d) breeding the modified female mouse of (c) with the modified male mouse of (c′) to obtain a mouse embryo homozygous for the genetic modification.
A method for making a mouse embryo heterozygous for a genetic modification, comprising:
- (a) incorporating a mouse ES donor cell into a pre-morula diploid mouse host embryo, wherein the mouse ES donor cell is heterozygous for the genetic modification and is from an inbred strain;
- (b) culturing the embryo to the blastocyst stage;
- (c) gestating the embryo of (b) in a surrogate mother, wherein upon gestation a modified mouse embryo is obtained having at least 90% cellular contribution from the mouse ES donor cell.
A method for making a mouse heterozygous for a genetic modification, comprising:
- (a) incorporating a mouse ES donor cell into a pre-morula diploid mouse host embryo, wherein the mouse ES donor cell is heterozygous for the genetic modification and is from an inbred strain;
- (b) culturing the embryo to the blastocyst stage;
- (c) gestating the embryo of (b) in a surrogate mother, wherein upon gestation a modified mouse is obtained having at least 90% cellular contribution from the mouse ES donor cell.
Dependencies are inferred from the claim text.