Methods and compositions for gene inactivation
Disclosed herein are methods and compositions for inactivating CCR-5 genes, using zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding ZFNs, vectors comprising polynucleotides encoding ZFNs, such as adenovirus (Ad) vectors, and cells comprising polynucleotides encoding ZFNs and/or cells comprising ZFNs are also provided.
1 . A method for inactivating an HIV receptor gene in a subject, the method comprising:
introducing, into a cell of the subject, a homing endonuclease that binds to a target site in an HIV receptor gene and cleaves the HIV receptor gene.
2 . The method of claim 1 , wherein the homing endonuclease is engineered to bind to the target site in the HIV receptor gene.
3 . The method of claim 1 , wherein a polynucleotide encoding the meganuclease is introduced into the cell.
4 . The method of claim 1 , wherein the homing endonuclease is introduced into the cell ex vivo.
5 . The method of claim 1 wherein the homing endonuclease is introduced into the cell in vivo.
6 . The method of claim 3 , wherein the homing endonuclease is introduced into the cell ex vivo.
7 . The method of claims 3 , wherein the homing endonuclease is introduced into the cell in vivo.
8 . The method of claim 1 , wherein the HIV receptor is a CCR-5 receptor.
9 . The method of claim 1 , wherein the HIV receptor is a CXCR4 receptor.
10 . The method of claim 2 wherein the cell is selected from the group consisting of a stem cell, a T-cell, a macrophage, a dendritic cell and an antigen-presenting cell.
11 . The method of claim 1 , wherein the homing endonuclease is selected from the group consisting of I-SceI, I-CeuI, PI-PspI, PI-Sce, I-SceIV, I-CsmI, I-PanI, I-SceII, I-PpoI, I-SceIII, I-CreI, I-TevI, I-TevII and I-TevIII.
12 . The method of claim 9 , wherein the homing endonuclease is SceI or CreI.
13 . A method of treating or preventing HIV infection in a subject, the method comprising:
inactivating an HIV receptor according to the method of claim 1 in a cell isolated from the subject; and
introducing the cell into the subject.
14 . A method of treating or preventing HIV infection in a subject, the method comprising:
introducing the polynucleotide encoding the meganuclease according to claim 7 into a subject; and
inactivating an HIV receptor.