# US 8,772,453 B2 · Compositions for linking zinc finger modules

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 8,772,453 B2
- Document ID: US8772453B2
- Status: Granted
- Kind: B2
- Application number: 13/068,102
- Filed: 2011-05-02
- Published: 2011-11-24
- Granted: 2014-07-08
- Earliest claimed: 2010-05-03
- Assignees: Sangamo BioSciences, Inc.
- Applicants: David Paschon; Edward J. Rebar
- Inventors: David Paschon; Edward J. Rebar
- Web page: https://patentgrep.com/patent/US8772453B2

## CPC

- C12N 15/62: DNA sequences coding for fusion proteins
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/04: Antibacterial agents
- A61P 31/12: Antivirals
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Disclosed herein are compositions for linking DNA binding modules to allow for specific and selective binding to module subsites separated by 1 or more base pairs. Also described are methods of making and using compositions comprising these linkers.

## Claims

### Claim 1

A zinc finger protein comprising a plurality of non-naturally occurring zinc finger modules, wherein at least two of the non-naturally occurring zinc finger DNA-binding modules are joined by an amino acid linker sequence of 5 to 20 amino acid residues between the last residue of the N-terminal zinc finger module and the first residue of C-terminal zinc finger module, the amino acid linker comprising an N-terminal amino acid residue adjacent to the N-terminal zinc finger module, a C-terminal amino acid residue adjacent to the C-terminal zinc finger module, and amino acid residues internal to the N- and C-terminal amino acid linker residues, wherein
(i) the N-terminal amino acid linker residue or the internal amino acid linker residues comprises at least one proline residue;
(ii) the zinc finger protein binds to a target site comprising target subsites bound by each zinc finger module; and
(iii) there is a gap of 0, 1, 2 or 3 base pairs between the target subsites bound by the zinc finger DNA-binding modules joined by the amino acid linker.

### Claim 2 (depends on 1)

The zinc finger protein of claim 1, wherein the linker further comprises at least two basic amino acid residues.

### Claim 3 (depends on 2)

The zinc finger protein of claim 2, wherein the basic amino acid residues are selected from the group consisting of an arginine residue, a histidine residue, a lysine residue or combinations thereof.

### Claim 4 (depends on 1)

The zinc finger protein of claim 1, wherein one or more of the zinc finger proteins further comprises a regulatory domain.

### Claim 5 (depends on 4)

The zinc finger protein of claim 4, wherein the regulatory domain is a transcriptional modulating domain.

### Claim 6 (depends on 5)

The zinc finger protein of claim 5, wherein the regulatory domain is an activation domain or a repression domain.

### Claim 7 (depends on 4)

The zinc finger protein of claim 4, wherein the regulatory domain is a cleavage domain or cleavage half-domain.

### Claim 8 (depends on 1)

An isolated polynucleotide encoding the zinc finger protein according to claim 1.

### Claim 9 (depends on 1)

An isolated cell comprising the zinc finger protein according to claim 1.

### Claim 10 (depends on 8)

An isolated cell comprising the polynucleotide according to claim 8.

### Claim 11 (depends on 1)

A method of modulating expression of a gene in a cell, the method comprising administering the zinc finger protein according to claim 1.

### Claim 12 (depends on 11)

The method of claim 11, wherein the zinc finger protein is administered to the cell as a polynucleotide.

### Claim 13 (depends on 11)

The method of claim 11, wherein the modulation comprises up or down regulation of the gene.

### Claim 14 (depends on 11)

The method of claim 11, wherein the modulation comprises cleavage of the gene.

### Claim 15 (depends on 11)

The method of claim 11, wherein the gene is an endogenous cellular gene.