Compositions and methods for the synthesis of fatty acids, their derivatives and downstream products

• US 20030159164A1
• Filed: 10/23/2002
• Published: 08/21/2003
• Est. Priority Date: 05/29/1998
• Status: Abandoned Application

• Current Assignee: Bruce Kelder, John Joseph Kopchick, Pradip Mukerji, Stephen J. Kirchner, Yung-Sheng Huang
• Original Assignee: Bruce Kelder, John Joseph Kopchick, Pradip Mukerji, Stephen J. Kirchner, Yung-Sheng Huang
• Inventors: Huang, Yung-Sheng, Kirchner, Stephen J., Mukerji, Pradip, Kelder, Bruce, Kopchick, John Joseph
• Application Number:
  US10/278,391
• Publication Number:
  US 20030159164A1
• Time in Patent Office: Days
• US Class Current: 800/14
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)
• CPC Class Codes: C12Y 114/19006 DELTA12-fatty-acid desaturase (1.14.19.6), i.e. oleoyl-CoA DELTA12 desaturase

Abstract

The invention generally relates to synthesis of essential fatty acids and their derivatives, long-chain polyunsaturated fatty acids (LC-PUFAs) and eicosanoids in transfected cells and in transgenic animals.

Claims

1. A composition, comprising animal cells producing essential fatty acids.

2. The composition of claim 1, wherein said animal cells comprise nucleic acid encoding a heterologous desaturase.

3. The composition of claim 1, wherein said nucleic acid comprises a heterologous desaturase gene selected from the group consisting of Δ

5-desaturase, Δ

6-desaturase, Δ

12-desaturase, and Δ

15-desaturase genes.

4. The composition of claim 3, wherein said animal cells are mammalian cells.

5. The composition of claim 4, wherein said animal cells produce arachidonic acid.

6. The composition of claim 4, wherein said mammalian cells are in tissue culture.

7. The composition of claim 4, wherein said mammalian cells are in a bioreactor.

8. A nutritional formulation, comprising at least one essential fatty acid derived from the animal cells of claim 1.

9. An animal feed formulation, comprising at least one essential fatty acid derived from the animal cells of claim 1.

10. A composition, comprising transfected animal cells, said transfected animal cells producing altered levels of long-chain polyunsaturated fatty acids relative to the animal cells prior to transfection.

11. The composition of claim 10, wherein said animal cells comprise nucleic acid encoding a heterologous desaturase.

12. The composition of claim 11, wherein said nucleic acid comprises a heterologous desaturase gene selected from the group consisting of Δ

5-desaturase, Δ

6-desaturase, Δ

12-desaturase, and Δ

15-desaturase genes.

13. The composition of claim 10, wherein said animal cells are mammalian cells.

14. The composition of claim 13, wherein said mammalian cells are in tissue culture.

15. The composition of claim 13, wherein said mammalian cells are in a bioreactor.

16. A nutritional formulation, comprising at least one long-chain polyunsaturated fatty acid derived from the animal cells of claim 10.

17. A cosmetic formulation, comprising at least one long-chain polyunsaturated fatty acid derived from the animal cells of claim 10.

18. An animal feed formulation, comprising at least one long-chain polyunsaturated fatty acid derived from the animal cells of claim 10.

19. A method, comprising:

a) providing;

i) a non-human animal cell, ii) a vector comprising nucleic acid encoding a heterologous desaturase, and iii) a recipient non-human female animal;

b) introducing said vector into said cell to create a transfected cell; and

c) transferring said transfected cell into said recipient female under conditions such that at least one offspring is produced, said offspring expressing said desaturase in one or more tissues.

20. The method of claim 19, wherein said offspring expresses said desaturase in said offspring'"'"'s mammary tissue, resulting in the production of essential fatty acids in the offspring'"'"'s milk.

21. The method of claim 20, further comprising step d) collecting said milk.

22. The method of claim 21, further comprising step e) isolating said essential fatty acids in said milk.

23. The method of claim 22, further comprising step (f) synthesizing one or more downstream products from said essential fatty acids islated in step (e).

24. The method of claim 23, wherein said one or more downstream products is selected from the group consisting of those downstream products set forth in FIGS. 12, 13, 14, 15, 16 and 17.

25. The method of claim 23, wherein said one or more downstream products is selected from the group consisting of leukotrienes and thromboxanes.

26. The method of claim 19, wherein said nucleic acid comprises a desaturase gene selected from the group consisting of Δ

5-desaturase, Δ

6-desaturase, Δ

12-desaturase, and Δ

5-desaturase genes.

27. The method of claim 26, wherein said desaturase gene is obtained from a fungal source.

28. The method of claim 26, wherein said desaturase gene is obtained from a plant source.

29. The method of claim 19, wherein said non-human female is selected from the group consisting of mice, rats, rabbits, pigs, goats, sheep, cows and horses.

30. A composition, comprising the milk of the offspring produced according to the method of claim 20.

31. A transgenic non-human animal, said animal producing at least one product, said product selected from the group consisting of arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid.