Use of non-subtype b gag proteins for lentiviral packaging

Abstract

The invention encompasses a lentiviral packaging vector comprising a non-subtype B gag-pol sequence, particularly a subtype D gag-pol sequence. The invention further encompasses methods for making and using these vectors. The invention further encompasses lentiviral vector particles comprising HIV-1 non-subtype B Gag and/or Pol proteins.

We claim: 
     
         1 . A replication-defective lentiviral packaging vector lacking a 4) site and encoding an HIV-1 Gag MA protein;
 wherein the HIV-1 Gag MA protein:   a) has an amino acid at position 12 that is not a glutamic acid,   b) has an amino acid at position 15 that is not an arginine,   c) does not have both a valine at position 46 and a leucine at position 61; and   wherein the lentiviral packaging vector encoding the HIV-1 Gag MA protein generates at least a 1.5 fold increase in the titer of a packaged lentiviral vector as compared to the lentiviral packaging vector encoding an HIV-1 Gag MA protein of HIV-1 BRU.   
     
     
         2 . The vector of  claim 1 , wherein the amino acid at position 12 is a lysine. 
     
     
         3 . The vector of  claim 1 , wherein the amino acid at position 12 is an arginine. 
     
     
         4 . The vector of  claim 1 , wherein the amino acid at position 15 is a threonine. 
     
     
         5 . The vector of  claim 1 , wherein the amino acid at position 15 is an alanine. 
     
     
         6 . The vector of  claim 1 , wherein the amino acid at position 46 is a leucine. 
     
     
         7 . The vector of  claim 1 , wherein the amino acid at position 61 is an isoleucine. 
     
     
         8 . The vector of  claim 1 , wherein the amino acid at position 61 is a methionine. 
     
     
         9 . The vector of  claim 2 , wherein the amino acid at position 15 is a threonine. 
     
     
         11 . The vector of  claim 2 , wherein the amino acid at position 46 is a leucine. 
     
     
         12 . The vector of  claim 2 , wherein the amino acid at position 61 is an isoleucine. 
     
     
         13 . The vector of  claim 9 , wherein the amino acid at position 46 is a leucine. 
     
     
         14 . The vector of  claim 9 , wherein the amino acid at position 61 is an isoleucine. 
     
     
         15 . The vector of  claim 13 , wherein the amino acid at position 61 is an isoleucine. 
     
     
         16 . The vector of  claim 15 , wherein the HIV-1 Gag MA protein is the HIV-1 Gag MA protein of HIV-1 NDK. 
     
     
         17 . The vector of  claim 16 , wherein the vector encodes the amino acid sequence of SEQ ID NO:4. 
     
     
         18 . A method for generating a packaging vector comprising inserting a Gag and/or Pol sequence from a non-subtype B HIV-1 virus into a plasmid under the control of a non-HIV promoter to generate the packaging vector. 
     
     
         19 . A method for generating a lentiviral vector comprising administering a packaging vector encoding a non-subtype B HIV-1 Gag or Pol protein to a cell with a lentiviral vector. 
     
     
         20 . The method of  claim 19 , further comprising selecting for a packaging vector that packages a higher titer of the lentiviral vector than a same packaging vector encoding a subtype B HIV-1 Gag or Pol protein.