# US 2019/0169640 A1 · Novel dna-binding proteins and uses thereof

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- Number: US 2019/0169640 A1
- Document ID: US20190169640A1
- Status: Published application
- Kind: A1
- Application number: 16/274,024
- Filed: 2019-02-12
- Published: 2019-06-06
- Earliest claimed: 2010-05-17
- Assignees: Sangamo Therapeutics, Inc.
- Applicants: Sangamo Therapeutics, Inc.
- Inventors: Philip D. Gregory; Jeffrey C. Miller; David Paschon; Edward J. Rebar; Siyuan Tan; Fyodor Urnov; Lei Zhang
- Web page: https://patentgrep.com/patent/US20190169640A1

## CPC

- C12N 15/907: {in mammalian cells}
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/02: Ophthalmic agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/195: from bacteria
- C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]}
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12N 15/62: DNA sequences coding for fusion proteins
- C12N 15/8213: {Targeted insertion of genes into the plant genome by homologous recombination}
- C12N 15/85: for animal cells
- C12Y 301/21004: Type II site-specific deoxyribonuclease (3.1.21.4)
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 2319/80: containing a DNA binding domain, e.g. Lacl or Tet-repressor
- 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
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12Y: ENZYMES
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

Disclosed herein are polypeptides, polynucleotides encoding, cells and organisms comprising novel DNA-binding domains, including TALE DNA-binding domains. Also disclosed are methods of using these novel DNA-binding domains for modulation of gene expression and/or genomic editing of endogenous cellular sequences.

## Claims

### Claim 1

An isolated polynucleotide encoding a non-naturally occurring TALE DNA-binding polypeptide comprising:
two or more TALE-repeat units, the TALE repeat units comprising a repeat variable di-residue (RVD); and wherein the N-terminal region lacks at least 152 amino acids of a full-length TALE protein and the C-terminal region is truncated to residue C+63, and further wherein the polypeptide binds to DNA.

### Claim 2 (depends on 1)

The polynucleotide of claim 1, wherein at least one TALE-repeat unit comprises an atypical repeat variable di-residue (RVD).

### Claim 3 (depends on 2)

The polynucleotide of claim 2, wherein the polypeptide comprises an atypical RVD as shown in Tables 27A or 27B.

### Claim 4 (depends on 1)

The polynucleotide of claim 1, wherein the N-terminal region of the polypeptide is truncated to residue N+134 to residue N+137.

### Claim 5 (depends on 1)

The polynucleotide of claim 1, wherein the C-cap of the polypeptide comprises a TALE repeat domain.

### Claim 6 (depends on 1)

The polynucleotide of claim 1, wherein the polypeptide further comprises a functional domain.

### Claim 7 (depends on 6)

The polynucleotide of claim 6, wherein the functional domain is a transcriptional activator or a transcriptional repressor.

### Claim 8 (depends on 1)

The polynucleotide of claim 1, wherein the functional domain comprises a nuclease domain.

### Claim 9 (depends on 8)

The polynucleotide of claim 8, wherein the nuclease domain is an endonuclease domain.

### Claim 10 (depends on 1)

A cell comprising the polynucleotide of claim 1.

### Claim 11 (depends on 1)

A pharmaceutical composition comprising the polynucleotide of claim 1.

### Claim 12 (depends on 10)

A pharmaceutical composition comprising a cell according to claim 10.