Compositions and method for regulation of transcription

• US 6,479,653 B1
• Filed: 07/13/2000
• Issued: 11/12/2002
• Est. Priority Date: 08/26/1997
• Status: Expired due to Term

• Current Assignee: Ariad Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: ARIAD Gene Therapeutics, Inc. (Takeda Pharmaceutical Company Limited)
• Inventors: Natesan, Sridaran, Gilman, Michael Z.
• Primary Examiner(s): Yucel, Remy
• Assistant Examiner(s): LOEB, BRONWEN
• Application Number:
  US09/615,917
• Time in Patent Office: 852 Days
• Field of Search: 435/320.1, 435/325, 536/23.4
• US Class Current: 536/23.4
• 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 14/005 from viruses
• CPC Class Codes: C07K 14/39 from yeasts
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/1055 Protein x Protein interaction, e.g. two hybrid selection
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 2710/16622 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The present invention relates to novel fusion proteins which activate transcription, to nucleic acid constructs encoding the proteins and their use in the genetic engineering of cells.

Claims

1. A recombinant nucleic acid encoding a fusion protein which comprises a tetramerization or tetramerization domain and a composite DNA binding domain.

2. The recombinant nucleic acid of claim 1 wherein said fusion protein comprises a tetramerization domain.

3. The recombinant nucleic acid of claim 1 wherein said fusion protein comprises a tetramerization domain.

4. The recombinant nucleic acid of claim 1 which further comprises a transcription activation domain.

5. The recombinant nucleic acid of claim 1 which further comprises a transcription repression domain.

6. The recombinant nucleic acid of claim 2 which further comprises a transcription activation domain.

7. The recombinant nucleic acid of claim 2 which further comprises a transcription repression domain.

8. The recombinant nucleic acid of claim 3, wherein the tetramerization domain is or is derived from a lac repressor tetramerization domain or a p53 tetramerization domain.

9. The recombinant nucleic acid of claim 3 which further comprises a transcription activation domain.

10. The recombinant nucleic acid of claim 3 which further comprises a transcription repression domain.

11. The recombinant nucleic acid of claim 8 which further comprises a transcription activation domain.

12. The recombinant nucleic acid of claim 8 which further comprises a transcription repression domain.

13. The recombinant nucleic acid of any one of claims 1, 2, 3, 8, 4, 6, 9, 11, 5, 7, 10 or 12 which further contains a ligand binding domain.

14. The recombinant nucleic acid of any one of claims 1, 2, 3, 8, 4, 6, 9, 11, 5, 7, 10 or 12, wherein the composite DNA binding domain comprises a homeodomain.

15. The recombinant nucleic acid of any one of claims 1, 2, 3, 8, 4, 6, 9, 11, 5, 7, 10 or 12, wherein the composite DNA binding domain comprises a zinc finger domain.

16. The recombinant nucleic acid of any one of claims 4, 6, 9 or 11 which comprises a transcription activation domain of 6 to 100 amino acids residues in length.

17. The recombinant nucleic acid of any one of claims 4, 6, 9 or 11 in which the transcription activation domain is or is derived from a VP16 or p65 transcription activation domain.

18. The recombinant nucleic acid any one of claims 5, 7, 10 and 12 in which the transcription repression domain is derived from a Krü

ppel domain.

19. The recombinant nucleic acid of claim 13 wherein the ligand binding domain is or is derived from an immunophilin, cyclophilin, FRB (FKBP:

rapamycin complex binding antibiotic binding domain or hormone receptor domain.

20. The recombinant nucleic acid of claim 16, wherein the composite DNA binding domain comprises a homeodomain.

21. The recombinant nucleic acid of claim 16, wherein the composite DNA binding domain comprises one or more zinc finger domains.

22. The recombinant nucleic acid of claim 17 which comprises part or all of the sequence corresponding to residues 361-550 of a human p65 protein.

23. The recombinant nucleic acid of claim 17, wherein the composite DNA binding domain comprises a homeodomain.

24. The recombinant nucleic acid of claim 17, wherein the composite DNA binding domain comprises one or more zinc finger domains.

25. The recombinant nucleic acid of claim 19 wherein the ligand binding domain is or is derived from FKBP (FK506 binding protein tetR tetracycline repressor progesterone receptor or ecdysone receptor.

26. The recombinant nucleic acid of claim 22, wherein the composite DNA binding domain comprises a homeodomain.

27. The recombinant nucleic acid of claim 22, wherein the composite DNA binding domain comprises one or more zinc finger domains.

28. A vector comprising a recombinant nucleic acid of any one of claims 1, 2, 3, 8, 4, 6, 9, 11, 5, 7, 10 or 12.

29. A vector comprising a recombinant nucleic acid of claim 14.

30. A vector comprising a recombinant nucleic acid of claim 15.

31. A vector comprising a recombinant nucleic acid of claim 20.

32. A vector comprising a recombinant nucleic acid of claim 21.

33. A vector comprising a recombinant nucleic acid of claim 23.

34. A vector comprising a recombinant nucleic acid of claim 24.

35. A vector comprising a recombinant nucleic acid of claim 26.

36. A vector comprising a recombinant nucleic acid of claim 27.

37. A cell comprising a recombinant nucleic acid of any one of claims 1, 2, 3, 8, 4, 6, 9, 11, 5, 7, 10 or 12.

38. A cell comprising a recombinant nucleic acid of claim 14.

39. A cell comprising a recombinant nucleic acid of claim 15.

40. A cell comprising a recombinant nucleic acid of claim 20.

41. A cell comprising a recombinant nucleic acid of claim 21.

42. A cell comprising a recombinant nucleic acid of claim 23.

43. A cell comprising a recombinant nucleic acid of claim 24.

44. A cell comprising a recombinant nucleic acid of claim 26.

45. A cell comprising a recombinant nucleic acid of claim 27.