Regulatory elements for delivery to the liver

Abstract

The invention is directed to novel combinations of liver specific enhancers and promoter elements for achieving persistent transgene expression in the liver. The liver specific enhancer elements may be derived from either the human serum albumin, prothrombin, α-1microglobulin or aldolase genes in single copies or in multimerized from linked to elements derived from the cytomegalovirus intermediate early (CMV), α-1-antitrypsin or albumin promoters. In a preferred embodiment of the invention, an adenoviral vector comprising a liver specific enhancer/promoter combination operably linked to a transgene is administered to recipient cells. In other embodiments of the invention, adeno-associated viral vectors, retroviral vectors, lentiviral vectors or a plasmid comprising the liver specific enhancer/promoter combination linked to a transgene is administered to recipient cells. Also within the scope of the invention are promoter elements derived from the human prothrombin gene and the β-fibrinogen gene.

1 . A recombinant transgene useful for expression of a coding sequence, comprising a strong constitutive promoter and one or more liver-specific enhancer elements.  
     
     
         2 . A recombinant transgene according to  claim 1 , wherein the strong constitutive promoter is selected from the group comprising a cytomegalovirus (CMV) promoter, a truncated CMV promoter, human serum albumin promoter and α-1-antitrypsin promoter.  
     
     
         3 . A recombinant transgene according to  claim 2 , wherein the promoter is a truncated CMV promoter from binding sites for known transcriptional repressors have been deleted.  
     
     
         4 . A recombinant transgene according to  claim 1 , wherein the liver-specific enhancer elements are selected from the group consisting of human serum albumin enhancers, human prothrombin enhancers, α-1microglobulin enhancers and intronic aldolase enhancers.  
     
     
         5 . A recombinant transgene according to  claim 1 , comprising one or more human serum albumin (HSA) enhancers and a promoter selected from the group consisting of a CMV promoter or an HSA promoter.  
     
     
         6 . A recombinant transgene according to  claim 1 , comprising one or more enhancer elements selected from the group consisting of human prothrombin (HPrT) enhancers and α-1microglobulin (A1MB) enhancers and the promoter is the CMV promoter.  
     
     
         7 . A recombinant transgene according to  claim 1 , wherein the enhancer elements are selected from the group consisting of HPrT enhancers and A1MB enhancers, and the promoter is the α-1-antitrypsin promoter.  
     
     
         8 . A recombinant adenoviral vector useful for transgene expression comprising a strong constitutive promoter and one or more liver-specific enhancer elements.  
     
     
         9 . A recombinant adenoviral vector according to  claim 8 , wherein the strong constitutive promoter is selected from the group comprising a cytomegalovirus (CMV) promoter, a truncated CMV promoter, human serum albumin promoter and α-1-antitrypsin promoter.  
     
     
         10 . A recombinant adenoviral vector according to  claim 9 , wherein the promoter is a truncated CMV promoter from binding sites for known transcriptional repressors have been deleted.  
     
     
         11 . A recombinant adenoviral vector according to  claim 8 , wherein the liver-specific enhancer elements are selected from the group consisting of human serum albumin enhancers, human prothrombin enhancers, α-1microglobulin enhancers and intronic aldolase enhancers.  
     
     
         12 . A recombinant adenoviral vector according to  claim 8 , comprising one or more human serum albumi (HSA) enhancers and a promoter selected from the group consisting of a CMV promoter or an HSA promoter.  
     
     
         13 . A recombinant adenoviral vector according to  claim 8 , comprising one or more enhancer elements selected from the group consisting of human prothrombin (HPrT) enhancers and α-1microglobulin (A1MB) enhancers and the promoter is the CMV promoter.  
     
     
         14 . A recombinant adenoviral vector according to  claim 8 , wherein the enhancer elements are selected from the group consisting of HPrT enhancers and A1MB enhancers, and the promoter is the α-1-antitrypsin promoter.