Methods and compositions for treating hemophilia B

• US 9,175,280 B2
• Filed: 10/12/2011
• Issued: 11/03/2015
• Est. Priority Date: 10/12/2010
• Status: Active Grant

• Current Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Original Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Inventors: Gregory, Philip D., High, Katherine A., Holmes, Michael C., Li, Hojun
• Primary Examiner(s): Babic, Christopher M
• Assistant Examiner(s): Hiriyanna, Kelaginamane T
• Application Number:
  US13/272,084
• Publication Number:
  US 20120128635A1
• Time in Patent Office: 1,483 Days
• Field of Search: 435/69.1
• US Class Current: 1/1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61N 2005/0661 ultraviolet
• CPC Class Codes: A61N 5/10 X-ray therapy; Gamma-ray therapy; Particle-irradiation therapy A61N5/01 takes precedence
• 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: C12N 15/1024 In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
• CPC Class Codes: C12N 9/644 Coagulation factor IXa (3.4.21.22)
• CPC Class Codes: C12Y 304/21022 Coagulation factor IXa (3.4.21.22)

Abstract

Disclosed herein are methods and compositions for insertion of Factor IX (FIX) sequences into the genome of a cell for treating hemophilia B.

Claims

1. A protein comprising an engineered zinc finger protein DNA-binding domain that binds to a target site in an endogenous Factor IX (FIX) gene, wherein the DNA-binding domain comprises four or five zinc finger recognition regions ordered F1 to F4 or F1 to F5 from N-terminus to C-terminus, and wherein

(i) when the DNA-binding domain comprises five zinc finger recognition regions, F1 to F5 comprise the following amino acid sequences:

2. The protein according to claim 1, further comprising a cleavage domain or cleavage half-domain.

3. The protein of claim 2, wherein the cleavage half-domain is a wild-type or engineered FokI cleavage half-domain.

4. A polynucleotide encoding the protein of claim 1.

5. A gene delivery vector comprising a polynucleotide according to claim 4.

6. An isolated cell comprising the protein of claim 1.

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

8. A method of integrating an exogenous sequence into an endogenous FIX gene in a cell, the method comprising:

expressing at least one polynucleotide according to claim 4 in the cell such that the exogenous sequence is integrated into the endogenous FIX gene.

9. A cell made by the method of claim 8.

10. The cell of claim 9, wherein the cell is a hepatic cell or a stem cell.

11. The cell of claim 10, wherein the stem cell is selected from the group consisting of an embryonic stem cell (ESC), an induced pluripotent stem cell (iPSC), a CD34+ hematopoietic stem cell and a hepatic stem cell.