Method of producing non-human mammals
First Claim
1. A method of producing a non-human mammal, referred to as an ES non-human mammal, wherein pluripotent cells are introduced into tetraploid blastocysts of the same mammalian species under conditions that result in production of an embryo and the resulting embryo is transferred into a foster mother which is maintained under conditions that result in development of live offspring, wherein the pluripotent cells are non-inbred pluripotent cells
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Abstract
A method of producing mutant/targeted non-human mammals, such as mutant mice that does not require production of chimera and permits the introduction of multiple mutations in embryos and, thus, avoids the necessity of breeding to combine all of the desired mutations in a single animal. The method is efficient in producing ES mice.
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Citations
48 Claims
- 1. A method of producing a non-human mammal, referred to as an ES non-human mammal, wherein pluripotent cells are introduced into tetraploid blastocysts of the same mammalian species under conditions that result in production of an embryo and the resulting embryo is transferred into a foster mother which is maintained under conditions that result in development of live offspring, wherein the pluripotent cells are non-inbred pluripotent cells
- 5. A method of producing a non-human mammalian embryo comprising injecting non-human non-inbred ES cells into non-human tetraploid blastocysts and maintaining the resulting tetraploid blastocysts under conditions that result in formation of embryos, thereby producing a non-human mammalian embryo.
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14. A method of producing a mutant non-human mammal, wherein pluripotent cells comprising at least one mutation in genomic DNA are introduced into tetraploid blastocysts of the same mammalian species under conditions that result in production of an embryo and the resulting embryo is transferred into a foster mother which is maintained under conditions that result in development of live offspring, thereby producing a mutant non-human mammal, wherein the pluripotent cells are non-inbred pluripotent cells.
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18. A method of producing a mutant non-human mammalian embryo comprising injecting mutant non-human non-inbred ES cells into non-human tetraploid blastocysts and maintaining the resulting tetraploid blastocysts under conditions that result in formation of embryos, thereby producing a mutant non-human mammalian embryo.
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27. A method of producing a mutant mouse, comprising:
- (a) introducing mouse non-inbred ES cells comprising at least one mutation in genomic DNA into mouse tetraploid blastocysts, thereby producing mouse blastocysts containing mouse non-inbred ES cells;
(b) maintaining the product of (a) under conditions that result in production of embryos;
(c) introducing an embryo into a pseudopregnant female; and
(d) maintaining the female into which the embryo is introduced under conditions that result in development of live offspring, thereby producing a mutant mouse.
- (a) introducing mouse non-inbred ES cells comprising at least one mutation in genomic DNA into mouse tetraploid blastocysts, thereby producing mouse blastocysts containing mouse non-inbred ES cells;
- 31. A mouse embryo produced from a mouse tetraploid blastocyst having incorporated therein mutant mouse non-inbred ES cells.
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35. A method of producing a mouse, comprising:
- (a) introducing mouse non-inbred ES cells into mouse tetraploid blastocysts, thereby producing mouse blastocysts containing mouse non-inbred ES cells;
(b) maintaining the product of (a) under conditions that result in production of embryos;
(c) introducing an embryo into a pseudopregnant female; and
(d) maintaining the female into which the embryo is introduced under conditions that result in development of live offspring, thereby producing a mouse.
- (a) introducing mouse non-inbred ES cells into mouse tetraploid blastocysts, thereby producing mouse blastocysts containing mouse non-inbred ES cells;
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38. A mouse embryo produced from a mouse tetraploid blastocyst having incorporated therein mouse non-inbred ES cells.
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41. A method of producing a mutant non-human mammal, mammal, wherein pluripotent cells comprising at least one mutation in genomic DNAare introduced into tetraploid blastocysts of the same mammalian species under conditions that result in production of an embryo and the resulting embryo is transferred into a foster mother which is maintained under conditions that result in development of live offspring, wherein the pluripotent cells are non-inbred pluripotent cells
- 42. A method of producing a mutant mouse that is derived from a single non-inbred ES cell clone, comprising breeding a mutant male mouse and a mutant female mouse, wherein the male mouse and the female mouse or an ancestor thereof were produced from the same non-inbred male ES cell and the female mouse is an XO female.
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44. A method of producing XO F1 ES cells, comprising introducing into male F1 ES cells a negative selection marker, under conditions appropriate for insertion of the negative selection marker in the Y chromosome of male F1 ES cells, thereby producing a mixture of male F1 Es cells comprising male F1 ES cells in which the negative selection marker in inserted in the Y chromosome and other male F1 ES cells, some of which do not contain a Y chromosome;
- subjecting the resulting mixture to conditions that result in the death of male F1 ES cells in which the Y chromosome has the negative selection marker inserted therein and do not result in the death of male F1 ES cells that lack a Y chromosome and are XO F1 ES cells, thereby producing XO F1 ES cells.
- View Dependent Claims (47, 48)
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45. An XO female mouse produced by introducing XO F1 ES cells into tetraploid mouse blastocysts under conditions that result in production of an embryo and transferring the resulting embryo into a foster mother which is maintained under conditions that result in development of live offspring, wherein the live offspring are XO female mice.
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46. A method of producing a mutant mouse strain, comprising breeding a mutant male mouse and a mutant female mouse, wherein the mutant male mouse and the mutant female mouse or an ancestor thereof were derived from the same non-inbred male mouse ES cell clone and the mutant female mouse is an XO female.
Specification