TALEN-based gene correction

Patent metadata

Field	Value
Publication number	US-9393257-B2
Application number	US-201414193037-A
Country	US
Kind code	B2
Filing date	Feb 28, 2014
Priority date	Mar 1, 2013
Publication date	Jul 19, 2016
Grant date	Jul 19, 2016

Abstract

The invention is directed to transcription activator-like effector nuclease (TALEN)-mediated DNA editing of disease-causing mutations in the context of the human genome and human cells to treat patients with compromised genetic disorders.

First claim

What is claimed is:

1. A composition comprising:
a nucleic acid encoding at least one TALEN protein, wherein the at least one TALEN protein is capable of inducing a site-specific double stranded DNA break in a target gene in a cell, wherein the target gene is a COL7A1 gene; and
a nucleic acid donor sequence,
wherein the donor sequence is a template for correction of a genetic mutation in the Col7A1 target gene, and further wherein the genetic mutation is capable of causing epidermolysis bullosa.

2. The composition of claim 1 , wherein the cell is selected from the group consisting of a fibroblast, keratinocyte, inducible pluripotent stem cell, hematopoietic stem cell, mesenchymal stem cell, embryonic stem cell, hematopoietic progeny cell, T-cell, B-cell, glial cell, neural cell, neuroglial progenitor cell, neuroglial stem cell, muscle cell, lung cell, pancreatic cell, liver cell and a cell of the reticular endothelial system.

3. The composition of claim 1 , wherein the composition comprises a nucleic acid encoding a first TALEN protein which is a left TALEN and the composition compromises a nucleic acid which encodes a second TALEN which is a right TALEN that cooperates with the left TALEN to make a site-specific double stranded DNA break in the target gene.

4. The composition of claim 1 , wherein the nucleic acid encoding the TALEN protein or the nucleic acid donor sequence is part of a vector or plasmid.

5. The composition of claim 3 , wherein the first TALEN and/or the second TALEN comprise a plurality of TAL effector repeat sequences and the endonuclease domain and a spacer between the plurality of TAL effector repeat sequences and the endonuclease domain includes a spacer.

6. The composition of claim 5 , wherein the spacer is 12 to 30 nucleotides in length.

7. A vector comprising the nucleic acid encoding the first TALEN protein of claim 1 .

8. A vector comprising the first nucleic acid and/or the second nucleic acid of claim 3 .

9. The composition of claim 1 , wherein the nucleic acid donor sequence comprises SEQ ID NO: 22.

10. The composition of claim 1 , wherein the nucleic acid sequence encoding the at least one TALEN protein is selected from the group consisting of SEQ ID Nos: 28, 29, 30, 31, and combinations thereof.

Assignees

• Univ Minnesota

Inventors

• Osborn Mark J
• Tolar Jakub
• Blazar Bruce
• Voytas Daniel F

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P7/06

  Antianaemics · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P19/08

  for bone diseases, e.g. rachitism, Paget's disease · CPC title

• A61P21/00

  Drugs for disorders of the muscular or neuromuscular system · CPC title