Vectors comprising SAR elements

Abstract

This invention relates to a method of using scaffolding attachment regions (SARs) to increase expression in retrovirally transduced resting cells. This includes gene therapy by introducing into a patient a cellular composition comprising non-immortal human cells transduced with a retroviral vector comprising a DNA SAR element and a heterologous gene operatively linked to an expression control sequence. A particularly preferred SAR is the 5′ SAR of the human interferon β gene or a fragment thereof.

1 . Use of a DNA scaffold or matrix attachment region (SAR) to increase gene expression in retrovirally transduced eukaryotic non-immortal resting cells.  
     
     
         2 . A method of modifying expression of a heterologous gene in a retrovirally transduced eukaryotic cell comprising transducing a non-immortal cell with a retrovirus comprising (i) at least one heterologous gene operatively linked to an expression control sequence and (ii) at least one SAR.  
     
     
         3 . A method of increasing expression of a heterologous gene in a retrovirally transduced eukaryotic resting cell comprising transducing a non-immortal eukaryotic resting cell with a retrovirus comprising (i) at least one heterologous gene operatively linked to an expression control sequence and (ii) at least one SAR, the SAR being in the reverse direction.  
     
     
         4 . A method of down regulating expression of a heterologous gene in a retrovirally transduced eukaryotic resting cell comprising transducing a non-immortal cell with a retrovirus comprising (i) at least one heterologous gene operatively linked to an expression control sequence and (ii) at least one SAR, the SAR being in the forward direction.  
     
     
         5 . A retroviral vector comprising genetic material corresponding to (a) at least one SAR, and (b) at least one heterologous gene operatively linked to an expression control sequence, the heterologous gene (or at least one of the heterologous genes if there is more than one heterologous gene) being rev-M10 and the SAR or at least one SAR is derived from, obtainable from or corresponds to the 5′ SAR of the human interferon-β gene.  
     
     
         6 . A cellular composition comprising non-immortal human cells transduced with a retroviral vector as claimed in  claim 5 .  
     
     
         7 . A cellular composition according to  claim 6  wherein the cells are hematopoietic cells.  
     
     
         8 . A cellular composition according to  claim 7  wherein the cells are T cells.  
     
     
         9 . A method of gene therapy for a patient in need thereof, comprising introducing into said patient, a cellular composition according to any one of claims  6 ,  7  or  8 .  
     
     
         10 . The method according to  claim 9  comprising the steps of removing hematopoietic cells from said patient, transducing said cells with a vector according to  claim 5 , and reintroducing the transduced cells into the patient.  
     
     
         11 . A method of treating a patient suffering from HIV infection, e.g., HIV-1 infection, comprising removing and isolating hematopoietic cells (e.g., hematopoietic stem cells, peripheral blood lymphocytes, CD4+ cells or T cells derived from hematopoietic stem cells) from said patient; transducing the cells with a gene for an anti-retroviral protein (e.g., rev-M10) and a SAR (e.g., a SAR derived or obtainable from the 5′ SAR of human IFN-β), and reintroducing the cells into the patient.  
     
     
         12 . All novel products, processes, and utilities substantially as described herein, especially with reference to the examples.