Assays for TERT promoter modulatory agents

Abstract

Methods and compositions for assaying an agent for TERT promoter modulatory activity are provided. In the subject methods, an agent is contacted with a cell comprising a mutant telomerase structural RNA component (TR) that results in a detectable phenotype in the presence of telomerase reverse transcriptase (TERT). Also provided are compositions, systems and kits thereof, as well as devices, that find use in practicing the subject methods. The subject invention finds use in assaying agents for TERT promoter modulatory activity, such as in a high throughput format.

1 . A method of determining whether an agent modulates transcription control activity of a TERT promoter, said method comprising:
 (a) contacting said agent with a cell comprising a mutant telomerase structural RNA component (TR) that results in a detectable phenotype in the presence of telomerase reverse transcriptase (TERT); and   (b) evaluating said cell for said detectable phenotype to determine whether said agent modulates transcription control activity of said TERT promoter nucleic acid.   
     
     
         2 . The method according to  claim 1 , wherein said TERT promoter nucleic acid is a human TERT promoter nucleic acid. 
     
     
         3 . The method according to  claim 1 , wherein said cell comprises an expression cassette that expresses said mutant telomerase structural RNA component (TR). 
     
     
         4 . The method according to  claim 3 , wherein said expression cassette is episomally maintained in said cell. 
     
     
         5 . The method according to  claim 3 , wherein said expression cassette is chromosomally integrated in said cell. 
     
     
         6 . The method according to  claim 5 , wherein said expression cassette is not chromosomally integrated into a chromosome of said cell that includes a TERT coding sequence. 
     
     
         7 . The method according to  claim 6 , wherein said cell is a human cell and said expression cassette is not integrated into chromosome 5. 
     
     
         8 . The method according to  claim 1 , wherein said detectable phenotype is cell death. 
     
     
         9 . The method according to  claim 1 , wherein said cell is a mutant cell that expresses telomerase and said method is a method for determining whether said agent inhibits expression controlled by a TERT promoter nucleic acid. 
     
     
         10 . The method according to  claim 1 , wherein said cell is a normal cell and said method is a method of determining whether said agent enhances expression controlled by a TERT promoter nucleic acid. 
     
     
         11 . The method according to  claim 1 , wherein said method comprises determining the modulatory activity of at least two different agents. 
     
     
         12 . The method according to  claim 11 , wherein said method is a high-throughput method. 
     
     
         13 . The method according to  claim 1 , wherein said agent is a small molecule. 
     
     
         14 . A method of determining whether a small molecule agent can derepress transcription repression activity of a TERT promoter, said method comprising:
 (a) contacting said agent with a cell comprising a mutant telomerase structural RNA component (TR) that results in a detectable phenotype in the presence of telomerase reverse transcriptase (TERT); and   (b) evaluating said cell for said detectable phenotype to determine whether said agent derepresses transcription repression activity of said TERT promoter nucleic acid.   
     
     
         15 . The method according to  claim 14 , wherein said cell is a human cell. 
     
     
         16 . The method according to  claim 15 , wherein said TERT promoter nucleic acid is a human TERT promoter nucleic acid. 
     
     
         17 . The method according to  claim 14 , wherein said cell comprises an expression cassette that expresses said mutant telomerase structural RNA component (TR). 
     
     
         18 . The method according to  claim 17 , wherein said expression cassette is episomally maintained in said cell. 
     
     
         19 . The method according to  claim 17 , wherein said expression cassette is chromosomally integrated in said cell. 
     
     
         20 . The method according to  claim 19 , wherein said expression cassette is not integrated into chromosome 5. 
     
     
         21 . The method according to  claim 14 , wherein said detectable phenotype is cell death. 
     
     
         22 . The method according to  claim 14 , wherein said method comprises determining the activity of at least two different agents. 
     
     
         23 . The method according to  claim 22 , wherein said method is a high-throughput method. 
     
     
         24 . The method according to  claim 14 , wherein said agent is a small molecule.