Alderon Biosciences proposes to develop a new electrochemical method for detection of specific nucleic acid sequences without enzymatic, fluorescent or radioactive labels and to show the feasibility of applying this method in an electrochemical plate (E-Plate) format to studies of gene expression in cancer. The E-Plate, an electrical analog of a traditional microplate and reader, will be developed as a tool to identify, quantify and study mRNAs and cDNA libraries of specific interest for cancer studies. Oligonucleotide probes containing electroactive bases will be used to capture the target sequences by hybridization on separately addressable elements of the E-Plate. Probe-target hybridization generates a signal in the presence of a buffer containing a mediator. The signal can be read out rapidly and be used to evaluate differential gene expression patterns. The expected result is an innovative electrochemical plate and reader that provides researchers with a rapid, facile and affordable method to probe small samples for specific mRNA or cDNA sequences. Another distinctive feature of the proposed approach that will be demonstrated in studies with human tumor suppressor mRNA targets is the ability to recover captured targets from the wells of the E-Plate for subsequent cloning, sequencing, or mass spectrometric analysis.
Thesaurus Terms: electrochemistry, gene expression, neoplasm /cancer genetics, nucleic acid quantitation /detection, technology /technique development electrode, guanine, nucleic acid hybridization, nucleic acid probe, oligonucleotide, oxidation reduction reaction, tumor suppressor gene mass spectrometry