Methods and compositions for prevention or treatment of a disease

• US 9,574,211 B2
• Filed: 05/13/2015
• Issued: 02/21/2017
• Est. Priority Date: 05/13/2014
• Status: Active Grant

• Current Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Original Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Inventors: Gregory, Philip D., Holmes, Michael C.
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US14/711,078
• Publication Number:
  US 20150329875A1
• Time in Patent Office: 650 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C07K 2319/71 containing domain for transcriptional activaation, e.g. VP16
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

Methods and compositions for treatment of a genetic disease are provided.

Claims

1. A fusion molecule comprising (i) an engineered nuclease that binds to a target site within SEQ NOs:

3, 4, 5 or 6 of an endogenous gene that produces a microRNA (miRNA), and (ii) a cleavage domain.

2. A polynucleotide encoding one or more fusion molecules of claim 1.

3. An isolated cell comprising one or more fusion molecules according to claim 1.

4. The cell of claim 3, wherein the cell is a stem cell.

5. The cell of claim 3, wherein the stem cell is selected from the group consisting of an embryonic stem cell (ESC), an induced pluripotent stem cell (iPSC), a hepatic stem cell and a liver stem cell.

6. The fusion molecule of claim 1, wherein the engineered nuclease is a zinc finger nuclease.

7. The zinc finger nuclease of claim 6 wherein the endogenous gene is MiR-34 and the zinc finger protein comprises 5 zinc, finger domains, each domain comprising a recognition helix region, wherein the zinc finger protein comprises the recognition helix regions as shown in a single row of Table III.

8. A kit comprising a fusion molecule according to claim 1.

9. A kit comprising a polynucleotide according to claim 2.

10. A method of cleaving an endogenous gene that produces an miRNA in a cell, the method comprising:

introducing, into the cell, one or more expression vectors comprising at least one polynucleotide according to claim 2, under conditions such that the one or more fusion molecules are expressed and the miRNA-producing gene is cleaved.

11. The method of claim 10, wherein the polynucleotide comprises an AAV vector.

12. The method of claim 10, wherein the polynucleotide is an mRNA.