METHODS AND COMPOSITIONS FOR TREATMENT OF A GENETIC CONDITION

• US 20180119175A1
• Filed: 12/19/2017
• Published: 05/03/2018
• Est. Priority Date: 05/15/2013
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Conway, Anthony, Cost, Gregory J., Dekelver, Russell, Rebar, Edward J., Reik, Andreas, Urnov, Fyodor, Wang, Jianbin, Zhang, H. Steve
• Granted Patent: US 10,196,651 B2
• Time in Patent Office: Days
• CPC Class Codes: C07K 14/805 Haemoglobins; Myoglobins
• CPC Class Codes: C07K 2/00 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 15/902 using homologous recombination
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Y 301/21004 Type II site-specific deoxyribonuclease (3.1.21.4)

Abstract

Methods and compositions for comprising single guide RNAs targeted to an endogenous gene for genetic alteration of cells are provided.

Claims

1. A CRISPR/Cas nuclease comprising a single guide RNA that binds to a target site in an enhancer region of a BCL11 a gene, wherein the nuclease cleaves and inactivates the BCL11a gene.

2. The nuclease of claim 1, comprising a Type IIS restriction enzyme nuclease domain or a Cas protein.

3. A mammalian cell comprising the nuclease of claim 1.

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

5. The mammalian cell of claim 4, wherein the stem cell is a hematopoietic stem cell.

6. A method of inactivating an endogenous BCL11A gene in a cell, the method comprising the steps of:

administering to the cell a CRISPR/Cas nuclease according to claim 1, wherein the nuclease cleaves and inactivates the BCL11a gene.

7. The method of claim 6, comprising a Type IIS restriction enzyme nuclease domain or a Cas protein.

8. The method of claim 6, wherein the cell is a mammalian cell.

9. The method of claim 8, wherein the mammalian cell is a stem cell.

10. The method of claim 9, wherein the stem cell is a hematopoietic stem cell.

11. A method of increasing expression of globin in a cell, the method comprising inactivating BCL11A expression in the cell according to the method of claim 6, such that globin expression is increased.

12. The method of claim 11, wherein the globin is fetal hemoglobin (HbF), β

-globin or γ

-globin.

13. A cell made by method of claim 6.

14. A method of treating a hemoglobinopathy in a subject in need thereof, the method comprising increasing globin expression in a cell of the subject according to the method of claim 13.

15. A method of treating a hemoglobinopathy in a subject in need thereof, the method comprising administering an isolated cell according to claim 14.

16. The method of claim 15, wherein the cell is stem cell.

17. The method of claim 16, wherein the stem cell is a hematopoietic stem cell.