DNA Based Folding Bio-Sensors That Are Capable of Detecting Transcription Factors Involved In Cancer Progression By Expressing Rcombinant TATA Binding Protein

Transcription factors are proteins that bind to a specific DNA sequence in the genome in order to control the flow of genetic information from DNA to mRNA.  This process, called transcription, is mediated by the RNA polymerase II complex, and ultimately dictates protein expression, and thus cellular function.  Transcription factors promote or block the recruitment of the RNA polymerase II complex to specific genes under different conditions, such as growth or response to disease.  One key transcription factor is TATA binding protein (TBP), which binds specifically to a sequence called the TATA box, a highly conserved region at the beginning of many genes. Thus, TBP binding to the TATA box helps position the RNA polymerase II complex over the transcription start site of the gene, allowing successful gene expression. Intriguingly, many varieties of cancer result from the aberrant regulation of transcription factors and other proteins that control and direct gene expression.  Therefore there is great interest, both academically and clinically, in a better understanding of how TBP and other transcription factors bind DNA and regulate these aberrant processes. Here, we report on the design and use of DNA-based folding bio-sensors that are capable of quantitatively detecting the transcription factors involved in cancer progression by expressing recombinant TBP and demonstrating sensitive detection using custom optical and electrochemical bio-sensors that are activated by TBP’s intrinsic DNA-binding activity.  These sensors will ultimately allow us to investigate the role that protein-protein interactions and inhibitory drugs have on gene expression.