In vivo delivery of nucleic acids to the liver or liver tissue

Abstract

The invention encompasses methods of delivering nucleic acids, including dsRNA, to mammalian target cells in vivo via intercellular transfer, wherein the dsRNA is delivered to or expressed in a first cell different from the target cell, wherein the first cell facilitates delivery of the dsRNA to the target cell.

What is claimed: 
     
         1 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to a target organ or tissue in an animal comprising:
 transfecting at least one cell of a first tissue in said animal with the nucleic acid or expression vector, wherein said transfection results in said nucleic acid or expression vector being delivered to at least one cell in said target tissue or organ, wherein said target organ or tissue is distal from the site of said transfected first cell.   
     
     
         2 . A nucleic acid or expression vector for use according to  claim 1 , wherein said first tissue is muscle and said at least one muscle cell is selected from the group consisting of myocytes and myoblasts. 
     
     
         3 . A nucleic acid or expression vector for use according to  claim 2 , wherein said nucleic acid or expression vector is expressed in transfected skeletal muscle cells. 
     
     
         4 . A nucleic acid or expression vector for use according to  claim 1 , wherein said first tissue is skin and said at least one skin cell is selected from cells in the dermal layer or subcutaneous layer of the skin. 
     
     
         5 . A nucleic acid or expression vector for use according to  claim 1 , wherein said nucleic acid or expression vector is a double-stranded RNA (dsRNA). 
     
     
         6 . A nucleic acid or expression vector for use according to  claim 5 , wherein said dsRNA corresponds to a target gene in said target organ or tissue and wherein delivery of said dsRNA to said organ or tissue inhibits or reduces expression of said target gene 
     
     
         7 . A nucleic acid or expression vector for use according to  claim 1 , wherein said transfected cells further express a transmembrane or surface ligand specific for a receptor on said target organ or tissue. 
     
     
         8 . A nucleic acid or expression vector for use according to  claim 7 , wherein said surface ligand is a lectin. 
     
     
         9 . A nucleic acid or expression vector for use according to  claim 7 , wherein said surface ligand is a galectin. 
     
     
         10 . A nucleic acid or expression vector for use according to  claim 7 ,
 wherein said transfection results in said nucleic acid or expression vector being delivered to at least one cell in said target tissue or organ expressing a lectin receptor.   
     
     
         11 . A nucleic acid or expression vector for use according to  claim 1 , wherein the nucleic acid is a dsRNA and wherein administration of said dsRNA to said at least one first cell does not trigger a detectable PKR response. 
     
     
         12 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to a target cell in an animal comprising: transfecting a first cell in vitro with said nucleic acid or expression vector, isolating at least one exovesicle from said first cell wherein said exovesicle contains said nucleic acid or expression vector, and contacting said target cell with said exovesicle. 
     
     
         13 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to a target organ or tissue in an animal wherein delivery comprises transfecting at least one first cell ex vivo with said nucleic acid or expression vector, and introducing said transfected cell into said animal, wherein said introduction results in production of an exovesicle containing said nucleic acid or expression vector and said nucleic acid or expression vector being delivered to at least one second cell in said target organ or tissue via said exovesicle, wherein said target organ or tissue is distal from the site of introduction of said transfected first cell. 
     
     
         14 . A nucleic acid or expression vector for use according to  claim 12 , wherein said first cell is a muscle cell or a skin cell. 
     
     
         15 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to a cancer cell in an animal comprising:
 transfecting at least one cell of a first tissue in said animal with the nucleic acid or expression vector, wherein said transfection results in production of an exovesicle containing said nucleic acid or expression vector and said nucleic acid or expression vector being delivered to at least one cancer cell via said exovesicle, wherein said cancer cell is distal from the site of said transfected first cell.   
     
     
         16 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to an HIV-infected cell in an animal comprising:
 transfecting at least one cell of a first tissue in said animal with the nucleic acid or expression vector, wherein said transfection results in said nucleic acid or expression vector being delivered to at least one HIV-infected cell, wherein said HIV-infected cell is distal from the site of said transfected first cell.   
     
     
         17 . A nucleic acid or expression vector for use in delivering a nucleic acid or expression vector to a target organ or tissue in an animal for treating a patient suffering from an infection comprising:
 transfecting at least one cell of a first tissue in said animal with the nucleic acid or expression vector, wherein said transfection results in said nucleic acid or expression vector being delivered to at least one cell in said target tissue or organ, wherein said target organ or tissue is distal from the site of said transfected first cell.   
     
     
         18 . A nucleic acid or expression vector for use according to  claim 17 , wherein said infection is by protozoa, bacteria, yeast, fungus or an intracellular parasite. 
     
     
         19 . A nucleic acid or expression vector for use according to  claim 15 , wherein said first tissue is muscle and said at least one muscle cell is selected from the group consisting of myocytes and myoblasts. 
     
     
         20 . A nucleic acid or expression vector for use according to  claim 15 , wherein said first tissue is skin and said at least one skin cell is selected from cells in the dermal layer or subcutaneous layer of the skin.