METHODS AND COMPOSITIONS FOR REGULATION OF TRANSGENE EXPRESSION

• US 20130177983A1
• Filed: 09/21/2012
• Published: 07/11/2013
• Est. Priority Date: 09/21/2011
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Rebar, Edward J.
• Granted Patent: US 9,394,545 B2
• Time in Patent Office: Days
• US Class Current: 435/375
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/04 Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2799/025 where the vector is derived from a parvovirus
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates

Abstract

Nucleases and methods of using these nucleases for expressing a transgene from a safe harbor locus in a secretory tissue, and clones and animals derived therefrom.

Claims

1. A non-naturally occurring fusion protein comprising a zinc finger protein that binds to an endogenous albumin gene and a cleavage domain, wherein the fusion protein modifies the endogenous albumin gene.

3. A polynucleotide encoding one or more fusion proteins of claim 1.

4. An isolated cell comprising one or more fusion proteins according to claim 1.

5. An isolated cell comprising one or more polynucleotides according to claim 3.

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

7. The cell of claim 6, 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.

8. The cell of claim 5, wherein the cell is a stem cell.

9. The cell of claim 8, 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.

10. A kit comprising a fusion protein according to claim 1.

11. A kit comprising a polynucleotide according to claim 3.

12. A method of cleaving an endogenous albumin gene in a cell, the method comprising:

introducing, into the cell, one or more expression vectors comprising at least one polynucleotide according to claim 3, under conditions such that the one or more fusion proteins are expressed and the albumin gene is cleaved.

13. The method of claim 12, wherein the polynucleotide comprises an AAV vector.

14. The method of claim 13, wherein the cell is a liver cell.

2. The fusion protein of claim 2, wherein the zinc finger protein comprises 4, 5 or 6 zinc finger domains comprising a recognition helix region, wherein the zinc finger proteins comprise the recognition helix regions shown in a single row of Table 1, Table 3, Table 5 or Table 8.