Nuclease-mediated regulation of gene expression
The present disclosure is in the field of genome engineering, particularly targeted modification of the genome of a hematopoietic cell.
1. An isolated genetically modified cell comprising a genomic modification at one cleavage site made by a zinc finger nuclease, (ZFN) or TAL-effector domain nuclease (TALEN), which ZFN or TALEN binds to a target site as shown in any of SEQ ID Nos:58-62, 65 or 74-77, wherein the genomic modification comprises insertions and/or deletions within the one cleavage site.
2. The genetically modified cell of claim 1 , wherein the cell is a stem cell.
3. The genetically modified cell of claim 2 , wherein the stem cell is a hematopoictic stem cell.
4. The genetically modified cell of claim 3 , wherein the hematopoietic stem cell is a CD34+ cell.
5. A genetically modified differentiated cell descended from the stem cell of claim 2 .
6. The genetically modified cell of claim 5 , wherein the cell is a red blood cell (RBC).
7. The genetically modified cell of claim 1 , wherein the nuclease is introduced into the cell as a polynucleotide.
8. The genetically modified cell of claim 1 , wherein the insertion comprises integration of a donor polynucleotide encoding a transgene.
9. A pharmaceutical composition comprising the genetically modified cell of claim 1 .
10. A DNA binding molecule comprising a TALE-effector protein (TALE) or a single guide RNA that binds to a target site as shown in any of SEQ ID Nos:58-62, 65 or 74-77.
11. A polynucleotide encoding one or more DNA-binding molecule of claim 10 .
12. An isolated cell comprising one or more DNA-binding molecules according to claim 10 .
13. A kit comprising a polynucleotide according to claim 11 .