Retroviral vectors

Abstract

Retroviral vector production systems for producing lentivirus-based vector particles which are capable of infecting and transducing non-dividing target cells, wherein one or more of the auxiliary genes such as vpr, vif, tat, and nef in the case of HIV-1 are absent, from the system. The systems and resulting retrovirus vector particles have improved safety over existing systems and vectors.

1 - 19 . (canceled)  
     
     
         20 . A lentivirus-based retroviral vector production system for producing a replication defective retroviral vector, wherein the retroviral vector production system comprises one or more nucleic acid sequences encoding a genome, gag, pol, and an envelope protein, wherein the retroviral vector production system lacks nucleic acid sequences encoding functional tat, and wherein the retroviral vector production system is capable of producing a replication defective retroviral vector.  
     
     
         21 . The retroviral vector production system according to  claim 20 , wherein tat is absent or disrupted in the vector system and is not functionally expressed in cells.  
     
     
         22 . The retroviral vector production system according to  claim 20 , further comprising a nucleic acid sequence encoding functionally active rev or RRE-type sequences.  
     
     
         23 . The retroviral vector production system according to  claim 22 , wherein at least one RRE-type sequence is a constitutive transport element (CTE).  
     
     
         24 . The retroviral vector production system according to  claim 23 , wherein the CTE is Mason Pfizer monkey virus CTE.  
     
     
         25 . The retroviral vector production system according to  claim 20 , further comprising at least one nucleotide sequence of interest (NOI).  
     
     
         26 . The retroviral vector production system according to  claim 25 , wherein the at least one NOI encodes a therapeutic protein or gene product of interest.  
     
     
         27 . A method for producing a replication defective retroviral vector comprising at least one NOI, comprising contacting the retroviral vector production system of  claim 25  with a cell, thereby producing the replication defective retroviral vector.  
     
     
         28 . An isolated cell comprising the retroviral vector production system of  claim 20 .  
     
     
         29 . A composition comprising the retroviral vector production system of  claim 20  and a carrier.  
     
     
         30 . The retroviral vector production system according to  claim 20 , wherein the nucleic acid sequences comprise DNA constructs which encode: (i) the genome, (ii) gag and pol proteins, and (iii) an envelope protein.  
     
     
         31 . The retroviral vector production system according to  claim 20 , wherein the genome comprises an operable promoter.  
     
     
         32 . The retroviral vector production system according to  claim 31 , wherein the promoter is a non-retroviral promoter.  
     
     
         33 . The retroviral vector production system according to  claim 20 , wherein the envelope protein is VSV-G.  
     
     
         34 . The retroviral vector production system according to  claim 20 , wherein the retroviral vector production system is based on HIV-1.  
     
     
         35 . A set of isolated nucleic acid sequences encoding the components of the retroviral vector production system according to  claim 20 , comprising a DNA construct which encodes the genome, a DNA construct which encodes gag and pol proteins, and a DNA construct which encodes an envelope protein.  
     
     
         36 . The set of nucleic acid sequences according to  claim 35 , further comprising a DNA construct which encodes a functionally active rev or RRE-type sequences.  
     
     
         37 . The set of nucleic acid sequences according to  claim 35 , wherein the DNA construct encoding the genome further comprises at least one NOI.  
     
     
         38 . A method for producing a replication defective retroviral vector, comprising expressing in a cell the retroviral vector production system according to  claim 20 , thereby producing the replication defective retroviral vector.