Zinc finger binding domains for GNN

• US 6,140,081 A
• Filed: 10/16/1998
• Issued: 10/31/2000
• Est. Priority Date: 10/16/1998
• Status: Expired due to Term

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Barbas, Carlos F.
• Primary Examiner(s): Carlson, Karen Cochrane
• Application Number:
  US09/173,941
• Time in Patent Office: 746 Days
• Field of Search: 530/350, 514/2, 536/23.1, 435/69.1, 435/320.1, 435/325, 435/252.3, 574/44
• US Class Current: 435/69.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters

Abstract

Zinc finger-nucleotide binding polypeptides having binding specificity for target nucleotides containing one or GNN triplets are provided. Compositions containing such polypeptides and the use of such polypeptides and compositions for regulating nucleotide function are also provided.

Claims

1. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

1.

2. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

2.

3. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

3.

4. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

4.

5. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

5.

6. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

6.

7. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

9.

8. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

10.

9. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

12.

10. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

13.

11. An isolated and purified zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

14.

12. An isolated and purified zinc finger-nucleotide binding polypeptide that contains from two to about 12 nucleotide binding regions, said nucleotide binding regions selected from the group consisting of the amino acid sequences depicted in SEQ ID NO:

1 through SEQ ID NO;

16.

13. The zinc finger-nucleotide binding polypeptide of claim 12, wherein said nucleotide binding polypeptide contains from two to about 6 nucleotide binding region, said nucleotide binding region selected from the group consisting of the amino acid sequence depicted in SEQ ID NO:

1 through SEQ ID NO;

16.

14. The zinc finger-nucleotide binding polypeptide of claim 12, wherein the nucleotide binding regions are operatively linked.

15. The zinc finger-nucleotide binding polypeptide of claim 14, wherein the nucleotide binding regions are operatively linked by a linker comprising the amino acid sequence of SEQ ID NO:

111.

16. The zinc finger-nucleotide binding polypeptide of claim 12, wherein each of the nucleotide binding regions bind to different nucleotide sequences.

17. The zinc finger-nucleotide binding polypeptide of claim 13, wherein said polypeptide binds to nucleotide sequence 5'"'"'-(GNN)_n -3'"'"', wherein each N is A, C, G, or T with the proviso that all N'"'"'s cannot be C and wherein n is 2 to 6.

18. The zinc finger-nucleotide binding polypeptide of claim 12 further operatively linked to one or more transcription regulating factors.

19. The zinc finger-nucleotide binding polypeptide of claim 13 further operatively linked to one or more transcription regulating factors.

37. An isolated and purified polynucleotide that encodes the zinc finger-nucleotide binding polypeptide of claim 12.

38. An isolated and purified polynucleotide that encodes the zinc finger-nucleotide binding polypeptide of claim 13.

39. An expression vector containing the polynucleotide of claim 37.

40. An expression vector containing the polynucleotide of claim 38.

41. A process of regulating a nucleotide sequence that contains the sequence (5'"'"'-GNN)n-3'"'"', where n is an integer from 1 to 6, the process comprising exposing the nucleotide sequence to an effective amount of the composition of claim 13.

42. The process of claim 41 wherein the sequence 5'"'"'-(GNN)n-3'"'"' is located in the transcribed region of the nucleotide sequence.

43. The process of claim 41 wherein the sequence 5'"'"'-(GNN)n-3'"'"' is located in a promotor region of the nuclcotide sequence.

44. The process of claim 41 wherein the sequence 5'"'"'-(GNN)n-3'"'"' is located within an expressed sequence tag.

45. The process of claim 41 wherein the composition is operatively linked to one or more transcription modulating factors.

20. A composition comprising from two to about 12 purified zinc finger-nucleotide binding polypeptides, wherein said polypeptides are selected from the group consisting of the amino acid sequences depicted in SEQ ID NO:

1 through SEQ ID NO;

16.

21. The composition of claim 20 containing from 2 to about 6 polypeptides.

22. The composition of claim 21 wherein the polypeptides are linked by a linker having the sequence of SEQ ID NO 111.

23. The composition of claim 21 that binds to a nucleotide that contains the sequence 5'"'"'-(GNN)_n -3'"'"', wherein each N is A, C, G, or T with the proviso that all N'"'"'s cannot be C and where n is 2 to 6.

24. The composition of claim 20 wherein the polypeptides are operatively linked.

25. The composition of claim 20 wherein each of the polypeptides binds to a different nucleotide sequence.

26. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

1.

27. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

2.

28. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

3.

29. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

4.

30. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

5.

31. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

6.

32. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

9.

33. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

10.

34. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

12.

35. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

13.

36. An isolated and purified polynucleotide that encodes a zinc finger-nucleotide binding polypeptide that contains the amino acid sequence depicted in SEQ ID NO:

14.