Bak promoter expression system

Abstract

The present invention provides polynucleotides which regulate the expression of a gene involved in apoptosis. Also provided are methods for identifying agents that modulate expression of a gene involved in apoptosis.

We claim:  
     
         1 . An isolated polynucleotide comprising the bak promoter (SEQ ID NO:1) or an active fragment thereof.  
     
     
         2 . The polynucleotide of  claim 1 , wherein said active fragment comprises positions −1077 to −1055 of said bak promoter.  
     
     
         3 . The polynucleotide of  claim 1 , wherein said active fragment comprises positions −2376 to −2360 of said bak promoter.  
     
     
         4 . The polynucleotide of  claim 1 , wherein said active fragment comprises positions −2984 to −2975 of said bak promoter.  
     
     
         5 . The polynucleotide of  claim 1 , wherein said active fragment comprises positions −3073 to −3064 of said bak promoter.  
     
     
         6 . The polynucleotide of  claim 1 , wherein said active fragment comprises positions −2591 to −2549 of said bak promoter.  
     
     
         7 . A method for identifying an agent that effectively regulates the expression of a gene involved in apoptosis in a cell, comprising the steps of: 
 a) introducing into said cell an isolated polynucleotide comprising a bak promoter or an active fragment thereof and a reporter gene;    b) determining the level of expression of said reporter gene in said cell of step (a);    c) contacting said cell of step (a) with the agent; and    d) identifying an effective agent that regulates the expression of said reporter gene.    
     
     
         8 . An effective agent, comprising the agent identified according to the method of  claim 7 .  
     
     
         9 . A method for identifying an agent that effectively increases the expression of a gene involved in apoptosis in a cell, comprising the steps of: 
 a) introducing into said cell an isolated polynucleotide comprising a bak promoter or an active fragment thereof and a reporter gene;    b) determining the level of expression of said reporter gene in said cell of step (a);    c) contacting said cell of step (a) with the agent; and    d) identifying an effective agent that increases the expression of said reporter gene.    
     
     
         10 . An effective agent, comprising the agent identified according to the method of  claim 9 .  
     
     
         11 . A method for identifying an agent that effectively decreases the expression of a gene involved in apoptosis in a cell, comprising the steps of: 
 a) introducing into said cell an isolated polynucleotide comprising a bak promoter or an active fragment thereof and a reporter gene;    b) determining the level of expression of said reporter gene in said cell of step (a);    c) contacting said cell of step (a) with the agent; and    d) identifying an effective agent that decreases the expression of said reporter gene.    
     
     
         12 . An effective agent, comprising the agent identified according to the method of  claim 11 .  
     
     
         13 . A method of reducing or inhibiting the level of apoptosis in a cell, comprising administering to the cell the effective agent of  claim 12 .  
     
     
         14 . A method of treating a patient having a disease characterized by an abnormal level of apoptosis, comprising administering to the patient a pharmaceutically acceptable composition comprising the effective agent of  claim 12 .  
     
     
         15 . A method of reducing or preventing toxicity of a normal cell in a patient receiving therapy, comprising administering to the patient a pharmaceutically acceptable composition comprising the effective agent of  claim 11 .  
     
     
         16 . The method of  claim 15 , wherein the therapy is chemotherapy.  
     
     
         17 . The method of  claim 15 , wherein the therapy is radiotherapy.