Regulation of endogenous gene expression in cells using zinc finger proteins
AssigneeSangamo BioSciences, Inc.
InventorsGeorge Norbert Cox, III, Casey Christopher Case, Stephen P. Eisenberg, Eric Edward Jarvis, Sharon Kaye Spratt
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Abstract
The present invention provides methods for modulating expression of endogenous cellular genes using recombinant zinc finger proteins.
Claims85
A method of inhibiting expression of an endogenous cellular pine in a cell, the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first engineered zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter,
- (ii) the nucleic acid molecule expresses the zinc finger protein is less cell;
- (iii) the zinc finger protein contacts a first target site in the endogenous cellular gene;
- and
- (iv) the Kd of the zinc finger protein is less than about 25 nM;
- thereby inhibiting expression of the endogenous cellular gene.
The method of claim wherein the step of administering further comprises administering a second zinc finger protein-encoding nucleic acid operably linked to a promoter that expresses a second zinc finger protein in the cell, and wherein the stop of contacting father comprises contacting a second target site in endogenous cellular gene with the second zinc finger protein.
A method of inhibiting expression of an endogenous cellular pine in a cell, the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first engineered zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter;
- (ii) the nucleic acid molecule expresses the zinc finger protein in the cell;
- (iii) the fusion zinc finger protein comprises six fingers and a regulatory domain;
- (iv) the fusion zinc finger protein contacts a target site in the endogenous cellular gene and;
- (v) the Kd of the zinc finger protein is less than about 25 nM;
- thereby inhibiting expression of the endogenous cellular gene.
A method of activating expression of an endogenous cellular gene in a cell the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first engineered zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter,
- (ii) the nucleic add molecule expresses a the zinc finger protein in the cell;
- (iii) the zinc finger protein contacts a first target site in the endogenous cellular gene;
- and
- (iv) the Kd of the zinc finger protein is less than about 25 nm;
- thereby activating expression of the endogenous cellular gene.
A method of activating expression of an endogenous cellular gene in a cell the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes an engineered fusion zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter;
- (ii) the nucleic acid molecule expresses the fusion zinc linger protein in the cell;
- (iii) the fusion zinc finger protein comprise six fingers and a regulatory domain;
- (iv) the fusion zinc finger protein contacts a target site in the endogenous cellular gene and;
- (v) the Kd of the fusion zinc finger protein is less than about 25 nm;
- thereby activating expression of the endogenous cellular gene.
A method of modulating expression of an endogenous cellular gene in a cell, the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first engineered zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter,
- (ii) the nucleic acid molecule expresses the zinc finger protein in the cell, and
- (iii) the zinc finger protein contacts a first target site in the endogenous cellular gene,
- thereby inhibiting expression of the endogenous cellular gene.
A method of modulating expression of an endogenous cellular gene in a cell, the method comprising the step of:
- administering to the cell a nucleic acid molecule comprising a polynucleic sequence which encodes an engineered fusion zinc finger protein, wherein
- (i) said polynucleotide sequence is operably linked to a promoter,
- (ii) the nucleic acid molecule, expresses the fusion zinc finger protein in the cell,
- (iii) the fusion zinc finger protein comprises six fingers and a regulatory domain, and
- (iv) the fusion zinc finger protein contacts a target site in the endogenous cellular gene;
- thereby modulating expression of the endogenous cellular gene.
Dependencies are inferred from the claim text.