Manipulation of negative stranded RNA viruses by rearrangement of their genes and uses thereof

Abstract

The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

What is claimed is:  
     
         1 . A method of constructing an attenuated virus useful for a vaccine, comprising the steps of: 
 rearranging gene order of said virus by moving a gene away from its wild-type 3′ promoter proximal position site, wherein said gene is an essential limiting factor for genome replication; and    placing a gene coding for an immune response-inducing antigen in the position closest to the 3′ end of the gene order of said virus, therefore, an attenuated virus is constructed for vaccine use.    
     
     
         2 . The method of  claim 1 , wherein the essential limiting factor gene is the nucleocapsid (N) gene.  
     
     
         3 . The method of  claim 1 , wherein the essential limiting factor gene is placed in the next to last position in the gene order of said virus.  
     
     
         4 . The method of  claim 1 , wherein the gene coding for an immune response-inducing antigen is selected from the group consisting of the attachment glycoprotein (G) gene, a fusion gene or the hemagglutinin/neuraminidase gene.  
     
     
         5 . A virus attenuated according to the method of  claim 1 .  
     
     
         6 . The method of  claim 1 , wherein said virus of the order Mononegavirales is a Rhabdovirus.  
     
     
         7 . The method of  claim 6 , wherein said Rhabdovirus is rabies virus or vesicular stomatitis virus.  
     
     
         8 . The method of  claim 1 , wherein said virus of the order Mononegavirales is a Paramyxovirus.  
     
     
         9 . The method of  claim 8 , wherein said Paramyxovirus is measles, mumps, parainfluenza virus or a respiratory syncytial virus.  
     
     
         10 . The method of  claim 9 , wherein said respiratory syncytial virus is a human respiratory syncytial virus or a bovine respiratory syncytial virus.  
     
     
         11 . The method of  claim 1 , wherein said virus of the order Mononegavirales is a Filovirus.  
     
     
         12 . The method of  claim 11 , wherein said Filovirus is Ebola virus or Marburg virus.  
     
     
         13 . The method of  claim 1 , wherein said attenuated virus useful for a vaccine is attenuated such that the lethal dose and the protective dose of the virus differ by about 1000 fold.