Adeno associated virus and adenovirus chimeric recombinant viruses useful for the integration of foreign genetic information into the chromosomal DNA of target cells

Abstract

Chimeric viral vectors having both the capacity to infect host cells efficiently and the capacity to integrate their genomic material into the host cell's genome. A chimeric viral vector having a functional packaging signal derived from a first virus and an integration means derived from a second virus. Typically, viruses capable of integrating their material into a host cell genome, having additional genetic material introduced therein recombinantly, do not have much room for insertion of such additional genetic material or are not very well capable of infecting every wanted host cell. Infecting viruses also lack a high insertion capacity or integration into the host cell's genome. The present invention provides integration of large inserts into a host cell's genome at an efficient infection rate by combining the properties of efficiently infecting viruses with efficiently integrating viruses.

What is claimed is:  
     
         1 . A chimeric viral vector comprising: 
 a functional packaging signal derived from a first virus having no integration means; and    an integration means derived from a second, integrating virus.    
     
     
         2 . The chimeric viral vector of  claim 1 , wherein the first virus is a virus capable of efficiently infecting a host.  
     
     
         3 . The chimeric viral vector of  claim 1  or  2 , wherein the first virus is an adenovirus.  
     
     
         4 . The chimeric viral vector of  claim 1 ,  claim 2 , or  claim 3  wherein the second, integrating virus is an adeno associated virus.  
     
     
         5 . The chimeric viral vector of  claim 4 , further comprising a sequence encoding a functional adeno associated virus rep gene.  
     
     
         6 . The chimeric viral vector of claiml,  claim 2 ,  claim 3 ,  claim 4  or  claim 5  further comprising a nucleic acid sequence of interest.  
     
     
         7 . The chimeric viral vector of  claim 6 , wherein the nucleic acid sequence of interest comprises one or more genes.  
     
     
         8 . The chimeric viral vector of  claim 6 , wherein the nucleic acid sequence of interest encodes a sequence chosen from a group consisting of antisense sequences and co-suppressing sequences.  
     
     
         9 . The chimeric viral vector of  claim 6 ,  claim 7 , or  claim 8  further comprising regulatory elements for expressing the nucleic acid sequence of interest.  
     
     
         10 . The chimeric viral vector of  claim 1 ,  claim 2 ,  claim 3 ,  claim 4 ,  claim 5 ,  claim 6 ,  claim 7 ,  claim 8 , or  claim 9  further comprising a suicide gene.  
     
     
         11 . A virus-like particle comprising a chimeric viral vector, the chimeric viral vector comprising: 
 a functional packaging signal derived from a first virus having no integration means; and    an integration means derived from a second, integrating virus.    
     
     
         12 . The virus-like particle of  claim 11 , further comprising an adenovirus-like particle.  
     
     
         13 . A method for producing a virus-like particle comprising a chimeric viral vector, the method comprising: 
 providing a cell with viral vector comprising 
 a functional packaging signal derived from a first virus having no integration means, and  
 an integration means derived from a second, integrating virus; and  
   providing said cell with elements necessary for producing said virus-like particle.    
     
     
         14 . The method according to  claim 13 , wherein a first element of said elements necessary for production of said virus-like particle is present in said vector and a second element of said elements necessary for production of said virus-like particle is present in a packaging vector present in said cell.  
     
     
         15 . The method according to  claim 13 , wherein said elements necessary for production of said virus-like particle are present on a packaging vector present in said cell.  
     
     
         16 . A packaging cell comprising genetic elements resulting in production of a virus-like particle including a chimeric viral vector having a functional packaging signal derived from a first virus lacking an integration means and an integration means derived from a second, integrating virus.  
     
     
         17 . The packaging cell of  claim 16  further comprising the vector according to any one of claims  1 - 10 .  
     
     
         18 . A cell comprising: 
 a chimeric viral vector comprising a functional packaging signal derived from a first virus lacking integration means and an integration means derived from a second, integrating virus.    
     
     
         19 . A virus-like particle according to  claim 11  or  12  for use as a pharmaceutical.  
     
     
         20 . A method for treatment of a host, said method comprising: 
 providing a host exhibiting a dysfunction selected from a group consisting of inherited diseases, malignancies, and genetic deficiencies;    providing a virus-like particle comprising a chimeric viral vector including a functional packaging signal derived from a first virus having no integration means and an integration means derived from a second, integrating virus; and    introducing said virus-like particle into said host.    
     
     
         21 . The virus-like particle of  claim 11  or  12  for use as a means for permanent genetic modification of a cell.  
     
     
         22 . The virus-like particle of  claim 11  or  12  for use as a means for making transgenic organisms.  
     
     
         23 . An rAAV nucleic acid comprising a nucleic acid sequence larger than 6.5 kb flanked by and physically linked with two ITRs, each of said two ITRs comprising an ITR selected from a group consisting of an AAV-ITR, a functional part of an AAV-ITR, and a derivative of an AAV-ITR.  
     
     
         24 . A method for the replication of an rAAV nucleic acid, the method comprising: 
 providing a cell with proteins encoded by at least a portion of an AAV-rep gene, at least a portion of a protein encoded by an adenovirus region selected from a group of adenovirus regions consisting of E1 regions, E2 regions, and E4 regions; and    providing said cell with a nucleic acid comprising an rAAV nucleic acid larger than 6.5 kb and being flanked by two AAV-ITR.    
     
     
         25 . The method according to  claim 24 , wherein providing said cell comprises providing a cell further comprising RNA encoded by at least a portion of an adenovirus VA-region.