Transgenic and cloned mammals

• US 20030177513A1
• Filed: 04/25/2003
• Published: 09/18/2003
• Est. Priority Date: 11/02/1998
• Status: Abandoned Application

• Current Assignee: GTC Biotherapeutics, Inc.
• Original Assignee: GTC Biotherapeutics, Inc.
• Inventors: Behbodi, Esmail, Echelard, Yann, Ziomek, Carol, Gavin, William, Melican, David
• Application Number:
  US10/423,792
• Publication Number:
  US 20030177513A1
• Time in Patent Office: Days
• US Class Current: 800/14
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/102 Caprine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0273 Cloned vertebrates
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/8121 Serpins
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/8772 Caprine embryos

Abstract

The invention features cloned and transgenic mammals, e.g., goats, which are made by introducing the genome of a somatic cell into an enucleated oocyte, preferably a naturally matured oocyte which in telophase, to form a reconstructed embryo.

Claims

1. A method of producing a cloned non-human mammal comprising:

introducing a genome from a non-human mammalian somatic cell into a naturally-matured oocyte which is in a telophase stage of meiotic cell division to form a reconstructed embryo; and

allowing the reconstructed embryo to develop into a mammal, thereby providing a non-human mammal.

2. The method of claim 1, wherein the mammal is a goat.

3. The method of claim 1, wherein the nucleus of the somatic cell is introduced into the oocyte.

4. The method of claim 1, wherein the somatic cell is a fibroblast.

5. The method of claim 4, wherein the fibroblast is an embryonic fibroblast.

6. The method of claim 1, wherein the somatic cell is in G₁ stage.

7. The method of claim 1, wherein the somatic cell is in G₀ stage.

8. The method of claim 1, wherein the oocyte is an enucleated oocyte.

9. The method of claim 3, wherein the nucleus of the somatic cell is introduced into the enucleated oocyte by fusion.

10. The method of claim 3, wherein the nucleus of the somatic cell is introduced into the enucleated oocyte by microinjection.

11. The method of claim 1, wherein the method further comprises, mating the mammal which develops from the reconstructed embryo with a second mammal.

12. A method of producing a transgenic non-human mammal, comprising:

introducing a genetically engineered genome of a mammalian somatic cell into a naturally matured oocyte which is in the telophase stage of meiotic cell division to form a reconstructed embryo; and

allowing the reconstructed embryo to develop into a mammal, thereby providing a transgenic mammal.

13. The method of claim 12, wherein the mammal is a goat.

14. The method of claim 12, wherein the nucleus of the genetically engineered somatic cell is introduced into the oocyte.

15. The method of claim 12, wherein the somatic cell is a fibroblast.

16. The method of claim 15, wherein the fibroblast is an embryonic fibroblast.

17. The method of claim 12, wherein the somatic cell is in G₁ stage.

18. The method of claim 12, wherein the somatic cell is in G₀ stage.

19. The method of claim 12, wherein the oocyte is an enucleated oocyte.

20. The method of claim 14, wherein the nucleus of the somatic cell is introduced into the enucleated oocyte by fusion.

21. The method of claim 14, wherein the nucleus of the somatic cell is introduced into the enucleated oocyte by microinjection.

22. The method of claim 12, wherein the method further comprises, mating the mammal which develops from the reconstructed embryo with a second mammal.

23. The method of claim 12, wherein the mammal is a female mammal.

24. The method of claim 23, wherein the mammal is induced to lactate.

25. The method of claim 12, wherein a product is recovered from the mammal.

26. The method of claim 25, wherein the product is recovered from the milk, urine, hair, blood, skin or meat of the mammal.

27. The method of claim 12, wherein the genome of the somatic cell comprises a transgenic sequence.

28. The method of claim 27, wherein the transgenic sequence encodes a human protein.

29. The method of claim 12, wherein the genome comprises a heterologous transgenic sequence under the control of a promoter.

30. The method of claim 29, wherein the promoter is a caprine promoter.

31. The method of claim 29, wherein the promoter is a tissue-specific promoter.

32. The method of claim 31, wherein the tissue specific promoter is a milk-specific promoter.

33. A method of making a cloned non-human mammal comprising:

fusing a mammalian somatic cell with a naturally matured enucleated oocyte which is in a telohase stage of meiotic division to obtain a reconstructed embryo;

activating the reconstructed embryo;

transferring the embryo into a doe; and

allowing the embryo to develop into a mammal.

34. The method of claim 33, wherein the mammal is a goat.

35. A method of making a transgenic non-human mammal comprising:

fusing a mammalian somatic cell capable of expressing a transgenic protein, with a naturally matured enucleated oocyte which is in a telophase stage of meiotic cell division to obtain a reconstructed embryo;

activating the reconstructed embryo;

transferring the embryo into a doe; and

allowing the embryo to develop into a mammal.

36. The method of claim 35, wherein the mammal is a goat.

37. The method of claim 35, wherein the nucleus of the somatic cell comprises a transgenic sequence.

38. The method of claim 35, wherein the transgenic sequence encodes a human protein.

39. A cloned non-human mammal, or descendant thereof, obtained by introducing a genome of a mammalian somatic cell into a naturally matured oocyte which is in a telophase stage of meiotic cell division to obtain reconstructed embryo and allowing the reconstructed embryo to develop into a mammal.

40. The mammal of claim 39, wherein the mammal is a goat.

41. The mammal of claim 39, wherein the genome is from a fibroblast.

42. The mammal of claim 41, wherein the fibroblast is an embryonic fibroblast.

43. A transgenic non-human mammal, or descendant thereof, obtained by introducing a genome of a genetically engineered mammalian somatic cell into a naturally matured oocyte which is in a telophase stage of meiotic cell division to obtain a reconstructed embryo and allowing the reconstructed embryo to develop into a mammal.

44. The mammal of claim 43, wherein the mammal is a goat.

45. The mammal of claim 43, wherein the genome of the genetically engineered somatic cell comprises a transgenic sequence.

46. The mammal of claim 45, wherein the transgenic sequence encodes a human protein.

47. The mammal of claim 43, wherein the genome comprises a heterologous transgenic sequence under the control of a promoter.

48. The mammal of claim 47, wherein the promoter is a tissue-specific promoter.

49. The mammal of claim 48, wherein the tissue specific promoter is a milk-specific promoter.