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 human hematopoietic stem or precursor cell comprising
(i) an endogenous BCL11a enhancer sequence; and
(ii) a pair of TALENs, each TALEN of the pair comprising a FokI cleavage half-domain and a DNA-binding domain comprising a plurality of TALE repeat units, each repeat unit comprising a hypervariable diresidue region (RVD), wherein the RVDs of the TALE repeats units are shown in a single row of as shown in Table 1, Table 2 or Table 4, and further wherein the pair of TALENs is selected from the group consisting of: 102740 and 102741; 102736 and 102737; 102756 and 102757; 102752 and 102753; 102750 and 102751; 102775 and 102774; 102795 and 102794; 102832 and 102833; 102834 and 102835; 102836 and 102837; 102838 and 102839; 102840 and 102841; 102842 and 102843; 102844 and 102845; 102846 and 102847; 102848 and 102849; 102850 and 102851; 102852 and 102853; 102854 and 102855; 102856 and 102857; 102858 and 102859; 102860 and 102861; 102862 and 102863; 102864 and 102865; 102866 and 102867; 102868 and 102869; 102870 and 102871; 102872 and 102873; 102874 and 102875; 102756 and 102757; 102752 and 102753; 102759 and 102751; 102876 and 102877; 102878 and 102879; 102880 and 102881; 102882 and 102883; 102884 and 102885; 102886 and 102887; 102888 and 102889; 102890 and 102891; 102892 and 102893; 102894 and 102895; 102896 and 102897; 102898 and 102899; 102902 and 102903; 102904 and 102905; 102906 and 102907; 102912 and 102913; 102914 and 102915; 102916 and 102917; 102918 and 102919; 102920 and 102921; or 102922 and 102923, wherein the pair of TALENs cleave and genetically modify the BCL11a enhancer sequence.
2. The cell of claim 1 , wherein the hematopoietic stem cell is a CD34+ cell.
3. The cell of claim 1 , wherein the TALEN pair is introduced into the cell as a polynucleotide.
4. A pharmaceutical composition comprising the cell of claim 1 .
5. A method of treating a human patient in need of an increase in gamma-globin gene or protein expression, the method comprising administering to the patient the pharmaceutical composition of claim 4 in an amount sufficient to increase the gamma-globin gene or protein expression in the human patient.
6. The method of claim 5 , wherein the human patient has β-thalassemia.