Methods and compositions for treatment of a genetic condition

• US 9,650,648 B2
• Filed: 08/29/2013
• Issued: 05/16/2017
• Est. Priority Date: 08/29/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Guschin, Dmitry, Cost, Gregory J., Gregory, Philip D., Reik, Andreas, Zhang, Lei, Holmes, Michael C., Miller, Jeffrey C., Paschon, David, Rebar, Edward J., Urnov, Fyodor
• Primary Examiner(s): SINGH, ANOOP KUMAR
• Application Number:
  US14/013,250
• Publication Number:
  US 20140080216A1
• Time in Patent Office: 1,356 Days
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/805 Haemoglobins; Myoglobins
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

Methods and compositions for a genetic disease are provided.

Claims

1. An isolated, non-naturally occurring fusion protein comprising a transcription activator like effector (TALE) DNA binding domain that binds to a target site in an endogenous BCL11A gene comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:

236, 238, 240 and 242, and a nuclease cleavage domain, wherein the fusion protein cleaves the endogenous BCL11A gene, wherein the TALE domain comprises a repeat variable diresidue (RVD) sequence in order presented and selected from a single row of table 1C.

2. An isolated polynucleotide encoding the non-naturally occurring fusion protein of claim 1.

3. An isolated cell comprising the non-naturally occurring fusion protein of claim 1.

4. An isolated cell comprising the polynucleotide of claim 2.

5. The cell of claim 4, wherein the cell is a red blood cell (RBC) precursor cell.

6. The cell of claim 3, wherein the cell is a red blood cell (RBC).

7. The cell of claim 5, wherein the RBC precursor cell is a CD34+hematopoietic stem cell.

8. A kit comprising the non-naturally occurring fusion protein of claim 1.

9. A kit comprising the polynucleotide of claim 2.

10. A method of increasing gamma-globin gene expression in an isolated cell, the method comprising:

transfecting the isolated cell with the polynucleotides of claim 2, wherein the isolated cell is a red blood cell (RBC) precursor cell or a hematopoietic stem cell, and

culturing said transfected cells under conditions such that said fusion protein is expressed and the endogenous BCL11A gene is cleaved, thereby increasing expression of the gamma-globin gene.

11. The method of claim 10, wherein the hematopoietic stem cell is a CD34+stem cell.