Compositions for linking DNA-binding domains and cleavage domains

• US 9,982,245 B2
• Filed: 07/11/2016
• Issued: 05/29/2018
• Est. Priority Date: 05/28/2008
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Miller, Jeffrey C.
• Primary Examiner(s): Hutson, Richard G
• Application Number:
  US15/206,768
• Publication Number:
  US 20160312197A1
• Time in Patent Office: 687 Days
• Field of Search: None
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/38 of the suprarenal hormones
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/80 containing a DNA binding domain, e.g. Lacl or Tet-repressor
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

Disclosed herein are compositions for linking DNA binding domains and cleavage domains (or cleavage half-domains) to form non-naturally occurring nucleases. Also described are methods of making and using compositions comprising these linkers.

Claims

1. A nuclease comprising:

a DNA-binding domain that binds to a nucleotide target site; and

a FokI cleavage domain, wherein the Fokl cleavage domain comprises a substitution of amino acid residues 161 and 162 of SEQ ID NO;

1 with an amino acid sequence EAAAR (SEQ ID NO;

5) or an insertion after amino acid residue 162 of SEQ ID NO;

1 of an amino acid sequence KSEAAAR (SEQ ID NO;

6).

2. The nuclease of claim 1, wherein the nucleotide target site is an endogenous gene.

3. A cell comprising the nuclease according to claim 1.

4. A kit for producing a nuclease, the kit comprising a nuclease according to claim 1 contained in one or more containers, optional hardware, and instructions for use of the kit.