Method of treating factor IX deficiency using nuclease-mediated targeted integration
Disclosed herein are methods and compositions for targeted, nuclease-mediated insertion of transgene sequences into the genome of a cell.
1. A method for treating a Factor IX (F.IX) deficiency in a mammal, the method comprising:
(i) intravenously administering one or more adeno-associated viral (AAV) vectors encoding a pair of zinc finger nucleases (ZFNs) or a pair of TAL-effector protein nucleases (TALENs) into a mammal with a F.IX deficiency such that a double-stranded break in an endogenous albumin gene occurs in liver cells of the mammal, and
(ii) intravenously administering one or more AAV vectors comprising a donor sequence comprising a transgene encoding a functional F.IX protein into the mammal 1 to 3 days after administering the one or more AAV vectors encoding the pair of ZFNs or TALENs, wherein the transgene is flanked by nucleic acid sequences homologous to the albumin gene, such that the transgene is integrated into the endogenous albumin gene, the functional F.IX protein is expressed in liver cells of the mammal at increased levels as compared to mammals in which the AAV vectors of (i) and (ii) are administered on the same day.
2. The method of claim 1 , wherein expression of the transgene is driven by an endogenous albumin promoter.
3. The method of claim 1 , wherein the F.IX protein is a is human F.IX (hF .IX) protein.