Methods and compositions for regulation of transgene expression

• US 9,777,281 B2
• Filed: 08/21/2015
• Issued: 10/03/2017
• Est. Priority Date: 09/21/2011
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Rebar, Edward J.
• Primary Examiner(s): KIM, ALEXANDER D
• Application Number:
  US14/832,434
• Publication Number:
  US 20150344893A1
• Time in Patent Office: 774 Days
• 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 polynucleotide encoding a non-naturally occurring fusion protein comprising

(i) a Transcription Activator Like Effector (TALE) DNA binding domain that binds to an endogenous albumin gene, wherein the TALE DNA binding protein comprises a plurality of TALE repeat units, each TALE repeat unit comprising an amino acid Repeat Variable Diresidue (RVD) that binds to a nucleotide in a target sequence in an endogenous albumin gene, wherein the TALE DNA binding domain comprises a +17 or +63C-terminal truncation and

(ii) a cleavage domain,

wherein the non-naturally occurring fusion protein cleaves the endogenous albumin gene.

2. An isolated cell comprising one or more polynucleotides according to claim 1.

3. The isolated cell of claim 2, wherein the cell is a stem cell.

4. 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.

5. A kit comprising the polynucleotide according to claim 1.

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

introducing, into the cell, one or more polynucleotides according to claim 1, under conditions such that the one or more fusion proteins—

are expressed and the endogenous albumin gene is cleaved.

7. The method of claim 6, wherein the one or more polynucleotides comprise mRNA.

8. The method of claim 6, wherein the one or more polynucleotides are included in one or more expression vectors.

9. The method of claim 6, wherein the cell is a liver cell.