Nuclease-mediated regulation of gene expression

Abstract

The present disclosure is in the field of genome engineering, particularly targeted modification of the genome of a hematopoietic cell.

1 - 19 . (canceled) 
     
     
         20 . A DNA-binding protein comprising a TALE-effector protein (TALE), wherein
 the TALE protein comprises 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 Table 1, Table 2 or Table 4.   
     
     
         21 . A fusion protein comprising a TALE protein of  claim 20  and a wild-type or engineered cleavage domain or cleavage half-domain. 
     
     
         22 . A polynucleotide encoding one or more proteins of  claim 20 . 
     
     
         23 . An isolated cell comprising one or more proteins according to  claim 20 . 
     
     
         24 . An isolated cell comprising one or more polynucleotides according to  claim 22 . 
     
     
         25 . The cell of  claim 23 , wherein the cell is a hematopoietic stem cell. 
     
     
         26 . A kit comprising a protein according to  claim 20 . 
     
     
         27 . A method of altering globin gene expression in a cell, the method comprising:
 introducing, into the cell, one or more polynucleotides according to  claim 22 , under conditions such that the one or more proteins are expressed and expression of the globin gene is altered.   
     
     
         28 . The method of  claim 27 , wherein expression of the globin gene is increased. 
     
     
         29 . The method of  claim 28 , wherein the globin gene is a gamma globin or beta globin gene. 
     
     
         30 . The method of  claim 27 , further comprising integrating a donor sequence into the genome of the cell. 
     
     
         31 . The method of  claim 30 , wherein the donor sequence is introduced to the cell using a viral vector, as an oligonucleotide or on a plasmid. 
     
     
         32 . The method of  claim 27 , wherein the cell is selected from the group consisting of a red blood cell (RBC) precursor cell and a hematopoietic stem cell. 
     
     
         33 . The method of  claim 30 , wherein the donor sequence comprises a transgene under the control of an endogenous or exogenous promoter. 
     
     
         34 - 39 . (canceled) 
     
     
         40 . A method of treating a patient in need of an increase in globin gene expression, the method comprising administering to the patient the pharmaceutical preparation of claim  19  in an amount sufficient to increase the globin gene expression in the patient. 
     
     
         41 . The method of  claim 40 , wherein the patient is known to have, is suspected of having, or is at risk of developing a globinopathy. 
     
     
         42 . The method of  claim 41 , wherein the globinopathy is a thalassemia or sickle cell disease. 
     
     
         43 . The method of  claim 42 , wherein the thalassemia is β-thalassemia. 
     
     
         44 . A pharmaceutical composition comprising the cell of  claim 23 . 
     
     
         45 . The cell of  claim 25 , wherein the hematopoietic stem cell is a CD34+ cell.