Targeting BCL11A distal regulatory elements for fetal hemoglobin reinduction
View Patent ↗Provided herein are methods and compositions for increasing fetal hemoglobin levels in a cell by disrupting BCL11A expression at the genomic level. Also provided herein are methods and compositions relating to the treatment of hemoglobinopathies by reinduction of fetal hemoglobin levels.
1. A method for producing a genetically modified human hematopoietic progenitor cell having decreased BCL11A mRNA and protein expression, the method comprising contacting an isolated human hematopoietic progenitor cell with a vector comprising a polynucleotide encoding a DNA-targeting endonuclease that cleaves genomic DNA of the cell on chromosome 2 between position 60,716,189 and 60,728,612 (according to UCSC Genome Browser hg 19 human genome assembly) in DNAse hypersensitive sites (DHSs) +62, +58, or +55, thereby causing a genetic modification therein and reducing the mRNA and the protein expression of BCL11A.
2. The method of claim 1 , wherein the isolated human hematopoietic progenitor is a cell of the erythroid lineage.
3. The method of claim 1 , wherein the genetic modification is a deletion.
4. The method of claim 3 , wherein the deletion removes the entire region between chromosome 2 location 60,716,189-60,728,612 or removes one or more of the DHSs in the region.
5. The method of claim 3 , wherein the deletion removes the entire region between chromosome 2 location 60,716,189-60,728,612.
6. The method of claim 3 , wherein the deletion disrupts DHS +62.
7. The method of claim 3 , wherein the deletion disrupts DHS +58.
8. The method of claim 3 , wherein the deletion disrupts DHS +55.
9. A method for producing a genetically modified human hematopoietic progenitor cell having decreased BCL11A mRNA and protein expression, the method comprising introducing an RNA encoding a DNA-targeting endonuclease into an isolated hematopoietic progenitor cell that cleaves genomic DNA of the cell on chromosome 2 between position 60,716,189 and 60,728,612 (according to UCSC Genome Browser hg 19 human genome assembly) in DHSs +62, +58, or +55, thereby causing a genetic modification therein and reducing the mRNA and the protein expression of BCL11A.
10. The method of claim 9 , wherein the isolated human hematopoietic progenitor is a cell of the erythroid lineage.
11. The method of claim 9 , wherein the genetic modification is a deletion.
12. The method of claim 11 , wherein the deletion removes the entire region between chromosome 2 location 60,716,189-60,728,612 or removes one or more of the DHSs in the region.
13. The method of claim 11 , wherein the deletion removes the entire region between chromosome 2 location 60,716,189-60,728,612.
14. The method of claim 11 , wherein the deletion disrupts DHS +62.
15. The method of claim 11 , wherein the deletion disrupts DHS +58.
16. The method of claim 11 , wherein the deletion disrupts DHS +55.