Modification of CXCR4 using engineered zinc finger proteins
Disclosed herein are methods and compositions for modulating activity of CXCR4 genes, for example using zinc finger transcription factors (ZF-TFs) or zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding ZF-TFs or ZFNs, vectors comprising polynucleotides encoding ZF-TFs or ZFNs and cells comprising polynucleotides encoding ZF-TFs or ZFNs and/or cells comprising ZF-TF or ZFNs are also provided.
1. An isolated genetically modified cell comprising a genomic modification within exon 2 of a CXCR4 gene, wherein the genomic modification is introduced using a nuclease that binds to a target site as shown in any of SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:16, SEQ ID NO:21 or SEQ ID NO:25, the genomic modification comprising a deletion, wherein the CXCR4 gene is inactivated and further wherein if the genomic modification further comprises an insertion, the insertion is a non-coding sequence of less than 3 base pairs.
2. The genetically modified cell of claim 1 , wherein the genetic modification is within an endogenous sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:16, SEQ ID NO:21 and SEQ ID NO:25.
3. The genetically modified cell of claim 1 , wherein the cell is selected from the group consisting of a hematopoietic stem cell, a T-cell, a macrophage, a dendritic cell and an antigen-presenting cell.
4. The genetically modified cell of claim 1 , wherein the modification is introduced using at least one zinc finger nuclease.
5. The genetically modified cell of claim 4 , wherein the nuclease is introduced into the cell as a polynucleotide.
6. The genetically modified cell of claim 4 , wherein the zinc finger nuclease comprises 4 or 5 zinc finger domains comprising a recognition helix and further wherein the zinc finger proteins comprise the recognition helix regions in the order shown in a single row of Table 1.
7. The genetically modified cell of claim 1 , wherein an endogenous CCR5 gene is inactivated in the cell.
8. The genetically modified cell of claim 1 , wherein the cell is resistant to HIV infection.
9. The genetically modified cell of claim 7 , wherein the cell is resistant to HIV infection.
10. A method for treating or preventing HIV infection in a subject, the method comprising:
introducing a cell according to claim 1 into the subject.
11. A method for treating or preventing HIV infection in a subject, the method comprising:
introducing a cell according to claim 7 into the subject.