Gene correction of SCID-related genes in hematopoietic stem and progenitor cells

Patent metadata

Field	Value
Publication number	US-9833479-B2
Application number	US-201715458258-A
Country	US
Kind code	B2
Filing date	Mar 14, 2017
Priority date	Jul 30, 2014
Publication date	Dec 5, 2017
Grant date	Dec 5, 2017

Abstract

The present disclosure is in the field of genome engineering, particularly targeted integration of a functional SCID-related genes (e.g., IL2RG, RAG1 and/or RAG2 gene) into the genome of a cell for provision of proteins lacking or deficient in SCID.

First claim

What is claimed is:

1. A host cell comprising an exogenous sequence integrated into intron 1 of an endogenous IL2RG gene or intron 1 or 2 of an endogenous RAG gene using a nuclease comprising a zinc finger protein, a TAL-effector domain or a single guide RNA (sgRNA) DNA-binding domain that binds to a sequence comprising a target site as shown in any of SEQ ID NOs:47-60 or 81-83 or a target site bound by any SEQ ID NO:85-95.

2. The host cell of claim 1 , wherein the nuclease comprises the zinc finger protein that comprises recognition helix regions as shown in a single row of Table 1.

3. The host cell of claim 1 , wherein the sgRNA comprises a DNA-binding guide RNA as shown in Table 5.

4. The host cell of claim 1 , wherein the TALE-effector domain comprises hypervariable diresidues (RVDs) as shown in in a single row of Table 3.

5. The host cell of claim 1 , wherein the exogenous sequence is selected from the group consisting of: a sequence encoding an IL2RG polypeptide integrated into intron 1 of an endogenous IL2RG gene; a sequence encoding a RAG polypeptide integrated into intron 1 or 2 of an endogenous RAG gene; and combinations thereof.

6. The host cell of claim 5 , wherein the exogenous sequence comprises a cDNA selected from the group consisting of a sequence comprising exons 2 through 8 of a wild type IL2RG gene; a sequence comprising a full-length IL2RG gene; a sequence comprising exon 3 of a wild type RAG gene and a sequence comprising a full-length RAG gene.

7. The host cell of claim 1 , wherein the cell is a hematopoietic stem cell or an induced pluripotent stem cell (iPSC).

8. A method of treating or preventing SCID or Omenn Syndrome in a subject, the method comprising administering a host cell according to claim 1 to the subject.

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

Assignees

• Sangamo Therapeutics Inc

Inventors

• Conway Anthony
• Cost Gregory J
• Holmes Michael C
• Urnov Fyodor

Classifications

• A61K35/545

  Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells · CPC title

• A61K48/005

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title

• C12N9/22 Primary

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• A61K38/1709

  from mammals · CPC title

• C12N15/907

  in mammalian cells · CPC title