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
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.
대표청구항▼
1. A method of producing a mouse, wherein mouse non-inbred ES cells are introduced into mouse tetraploid blastocysts by injection under conditions that result in production of an embryo and the resulting embryo is transferred into a pseudopregnant female mouse which is maintained under conditions th
1. A method of producing a mouse, wherein mouse non-inbred ES cells are introduced into mouse tetraploid blastocysts by injection under conditions that result in production of an embryo and the resulting embryo is transferred into a pseudopregnant female mouse which is maintained under conditions that result in development of live offspring. 2. The method of claim 1, wherein injection is piezo microinjection. 3. A method of producing a mouse embryo comprising injecting mouse non-inbred ES cells into mouse tetraploid blastocysts and maintaining the resulting tetraploid blastocysts under conditions that result in formation of embryos, thereby producing a mouse embryo. 4. The method of claim 3, wherein the mouse non-inbred ES cells are mutant mouse non-inbred ES cells and are injected into mouse tetraploid blastocysts by piezo microinjection. 5. A method of producing a mutant mouse, wherein mouse non-inbred pluripotent ES cells comprising at least one mutation in genomic DNA are introduced into mouse tetraploid blastocysts by injection 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 mouse, wherein said foster mother is a mouse. 6. The method of claim 5, wherein injection is piezo microinjection. 7. A method of producing a mutant mouse embryo comprising injecting mutant mouse non-inbred ES cells into mouse tetraploid blastocysts and maintaining the resulting tetraploid blastocysts under conditions that result in formation of embryos, thereby producing a mutant mouse embryo. 8. The method of claim 7, wherein mutant mouse non-inbred ES cells are injected into mouse tetraploid blastocysts by piezo microinjection. 9. 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 by injection, 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 mouse; and (d) maintaining the female mouse into which the embryo is introduced under conditions that result in development of live offspring, thereby producing a mutant mouse. 10. The method of claim 9, wherein injection is piezo microinjection. 11. The method of claim 10, wherein the at least one mutation in genomic DNA is a gene knockout or exogenous DNA incorporated into the genomic DNA. 12. 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 tile female mouse or an ancestor thereof were produced from the same non-inbred male ES cell clone and the female mouse is an XO female. 13. The method of claim 12, wherein the non-inbred ES cell clone is a non-inbred F1 cell clone. 14. A method of producing mouse XO F1 ES cells, comprising introducing into mouse male F1 ES cells a negative selection marker, under conditions appropriate for insertion of the negative selection marker in the Y chromosome of mouse F1 ES cells, thereby producing a mixture of mouse F1 ES cells comprising F1 ES cells in which the negative selection marker is inserted in the Y chromosome and other F1 ES cells, some of which do not contain a Y chromosome; subjecting the resulting mixture to conditions that result in the death of F1 ES cells in which the Y chromosome has the negative not result in the death of F1 ES cells that lack a Y chromosome and are XO F1 ES cells, thereby producing mouse XO F1 ES cells. 15. 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 mo use ES cell clone and the mutant female mouse is an XO female. 16. The method of claim 15, wherein the non-inbred male mouse ES cell clone is an F1 male mouse ES cell. 17. The method of claim 16, wherein the mutant XO female mouse or an ancestor thereof was derived from a male mouse F1 ES cell by knocking out the Y chromosome of the F1 ES cell, thereby producing an XO F1 ES cell; introducing the XO F1 ES cell into a tetraploid mouse blastocyst 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, thereby producing an XO female offspring.
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이 특허에 인용된 특허 (2)
Erwin Wagner AT; Zhao-Qi Wang FR, Method for developing transgenic mice.
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