Zinc finger proteins for DNA binding and gene regulation in plants

• US 7,262,054 B2
• Filed: 01/22/2002
• Issued: 08/28/2007
• Est. Priority Date: 01/22/2002
• Status: Expired due to Term

• Current Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Original Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Inventors: Jamieson, Andrew, Li, Guofu
• Primary Examiner(s): COLLINS, CYNTHIA E
• Application Number:
  US10/470,180
• Publication Number:
  US 20040128717A1
• Time in Patent Office: 2,044 Days
• Field of Search: 800/295, 800/278, 800/288, 800/306, 435/419, 435/410, 435/468, 530/300, 530/350, 536/23.1, 536/23.4
• US Class Current: 435/410
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8217 Gene switch

Abstract

Disclosed herein are modified plant zinc finger proteins; compositions comprising modified plant zinc finger proteins and methods of making and using modified plant zinc finger proteins. The modified plant zinc finger proteins, in contrast to naturally-occurring plant zinc finger proteins, have a binding specificity that is determined by tandem arrays of modular zinc finger binding units.

Claims

1. A method for modulating gene expression in a plant cell;

the method comprising contacting the cell with a plant zinc finger protein (ZFP) comprising a tandem array of a plurality of zinc fingers, the plant ZFP having a non-plant ZFP structure and being engineered to bind to a target sequence, wherein the plant ZFP is obtained by assembling sequences from a naturally occurring plant ZFP in a non-plant structure by (i) deleting one or more amino acid residues between the adjacent zinc fingers of the naturally-occurring plant ZFP such that there are between about 5 and 50 amino acids between adjacent zinc fingers of the plant ZFP and (ii) engineering the plant ZFP to bind to the target sequence.

2. The method of claim 1, wherein the protein is expressed in the cell.

3. The method of claim 2, wherein a nucleic acid encoding the protein is introduced into the cell.

4. The method of claim 1, wherein the ZFP is engineered to bind to the target sequence by rational design.

5. The method of claim 1, wherein the ZFP is engineered to bind to the target sequence by selection.

6. The method of claim 5, wherein selection is accomplished through the use of a method selected from the group consisting of phage display, interaction trap, ribosome display and RNA-peptide fusion.