Methods and compositions for diagnosing disease

Abstract

The present invention relates to methods and compositions for diagnosing a disease or disorder in a subject by introducing into cells of the subject a diagnostic gene switch construct and monitoring expression of a reporter gene. The invention further relates to methods and compositions for monitoring the progression of a disease or disorder or the effectiveness of a treatment for a disease or disorder.

What is claimed is: 
     
         1 . A method of monitoring the level of a factor that is being administered to a subject for treatment for a disease or disorder, comprising:
 administering said treatment to said subject;   introducing into cells of said subject (1) a polynucleotide encoding a gene switch, said gene switch comprising at least one transcription factor sequence, wherein said at least one transcription factor sequence encodes a ligand-dependent transcription factor, linked to a diagnostic switch promoter, wherein the activity of the promoter is modulated by said factor that is being administered for treatment, and (2) a polynucleotide encoding a reporter gene linked to a promoter which is activated by said ligand-dependent transcription factor, to produce modified cells;   administering ligand to said modified cells; and   detecting reporter gene expression;   wherein the level of expression of said reporter gene indicates the level of the factor being administered for treatment.   
     
     
         2 . The method of  claim 1 , wherein said polynucleotide is introduced into said cells prior to the said treatment is administered to said subject. 
     
     
         3 . The method of  claim 2 , wherein a baseline level of reporter gene expression is determined prior to the administration of said treatment to said subject. 
     
     
         4 . The method of  claim 1 , further comprising the step of modifying the amount of factor being administered based on the level of reporter gene expression detected. 
     
     
         5 . The method of  claim 1 , wherein said polynucleotides are introduced into cells that have been isolated from said subject to produce modified cells, and the modified cells are re-introduced into said subject. 
     
     
         6 . The method of  claim 1 , wherein said method is carried out in vivo. 
     
     
         7 . The method of  claim 1 , wherein said gene switch is an EcR-based gene switch. 
     
     
         8 . The method of  claim 7 , wherein said ligand binds to the EcR ligand binding domain. 
     
     
         9 . The method of  claim 8 , wherein said ligand is a diacylhydrazine. 
     
     
         10 . The method of  claim 9 , wherein said ligand is selected from the group consisting of RG-115819, RG-115830, and RG-115932. 
     
     
         11 . The method of  claim 1 , wherein said gene switch comprises a first transcription factor sequence under the control of a first diagnostic switch promoter and a second transcription factor sequence under the control of a second diagnostic switch promoter, wherein the proteins encoded by said first transcription factor sequence and said second transcription factor sequence interact to form a protein complex which functions as a ligand-dependent transcription factor.