Methods and compositions for treatment of a genetic condition
Methods and compositions for a genetic disease are provided.
1. An isolated, non-naturally occurring fusion protein comprising a transcription activator like effector (TALE) DNA binding domain that binds to a target site in an endogenous BCL11A gene comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 236, 238, 240 and 242, and a nuclease cleavage domain, wherein the fusion protein cleaves the endogenous BCL11A gene, wherein the TALE domain comprises a repeat variable diresidue (RVD) sequence in order presented and selected from a single row of table 1C.
2. An isolated polynucleotide encoding the non-naturally occurring fusion protein of claim 1 .
3. An isolated cell comprising the non-naturally occurring fusion protein of claim 1 .
4. An isolated cell comprising the polynucleotide of claim 2 .
5. The cell of claim 4 , wherein the cell is a red blood cell (RBC) precursor cell.
6. The cell of claim 3 , wherein the cell is a red blood cell (RBC).
7. The cell of claim 5 , wherein the RBC precursor cell is a CD34+hematopoietic stem cell.
8. A kit comprising the non-naturally occurring fusion protein of claim 1 .
9. A kit comprising the polynucleotide of claim 2 .
10. A method of increasing gamma-globin gene expression in an isolated cell, the method comprising:
transfecting the isolated cell with the polynucleotides of claim 2 , wherein the isolated cell is a red blood cell (RBC) precursor cell or a hematopoietic stem cell, and
culturing said transfected cells under conditions such that said fusion protein is expressed and the endogenous BCL11A gene is cleaved, thereby increasing expression of the gamma-globin gene.
11. The method of claim 10 , wherein the hematopoietic stem cell is a CD34+stem cell.