Compositions and methods for protein production

• US 20060063231A1
• Filed: 09/08/2005
• Published: 03/23/2006
• Est. Priority Date: 09/16/2004
• Status: Abandoned Application

• Current Assignee: Sangamo Biosciences, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Jamieson, Andrew, Collingwood, Trevor, Bartsevich, Victor, Li, Xiao-Yong
• Application Number:
  US11/221,683
• Publication Number:
  US 20060063231A1
• Time in Patent Office: Days
• US Class Current: 435/69.100
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C07K 2319/09 containing a nuclear localisation signal
• CPC Class Codes: C07K 2319/43 containing a FLAG-tag
• CPC Class Codes: C07K 2319/71 containing domain for transcriptional activaation, e.g. VP16
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 2830/002 inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor

Abstract

Disclosed herein are methods and compositions for enhanced protein production and overexpression using engineered zinc finger proteins.

Claims

1. A method for regulating the transcription of a nucleotide sequence in a cell, the method comprising:

expressing, in the cell, a protein comprising SEQ ID NO;

7 wherein the protein binds to a target site;

wherein the target site is operatively linked to the nucleotide sequence.

2. The method of claim 1, wherein the target site comprises SEQ ID NO:

1.

3. The method of claim 2, wherein a plurality of target sites are operatively linked to the nucleotide sequence.

4. The method of claim 1, wherein the nucleotide sequence comprises a cDNA sequence.

5. The method of claim 1, wherein the protein further comprises a transcriptional activation domain.

6. The method of claim 5, wherein the transcriptional activation domain is the VP16 domain.

7. A method for regulating the transcription of first and second nucleotide sequences in a cell, the method comprising:

expressing, in the cell, a protein comprising SEQ ID NO;

7 wherein the protein binds to a target site;

wherein the target site is operatively linked to both of the first and second nucleotide sequences.

8. The method of claim 7, wherein the target site comprises SEQ ID NO:

1.

9. The method of claim 8, wherein a plurality of target sites are operatively linked to the first nucleotide sequence.

10. The method of claim 8, wherein a plurality of target sites are operatively linked to the second nucleotide sequence.

11. The method of claim 8, wherein a plurality of target sites are operatively linked to both of the first and second nucleotide sequences.

12. The method of claim 7, wherein the first nucleotide sequence comprises a cDNA sequence.

13. The method of claim 7, wherein the second nucleotide sequence comprises a cDNA sequence.

14. The method of claim 7, wherein both of the first and second nucleotide sequences comprise cDNA sequences.

15. The method of claim 14, wherein the cDNA sequences encode antibody polypeptides.

16. The method of claim 15 wherein the first cDNA sequence encodes an antibody heavy chain and the second cDNA sequence encodes an antibody light chain.

17. The method of claim 7, wherein the protein further comprises a transcriptional activation domain.

18. The method of claim 17, wherein the transcriptional activation domain is the VP16 domain.