SIGNAL AMPLIFICATION FOR SPECIFIC NUCLEIC ACID DETECTION

THIS PROJECT WILL ATTEMPT TO COMMERCIALIZE A NOVEL, HIGH-SPEED, AND AUTOMATABLE DNA AND RNA DIAGNOSTIC TECHNOLOGY FOR THE SPECIFIC DETECTION OF NUCLEIC ACID SEQUENCE CHANGES ASSOCIATED WITH GENETIC DISEASES. THIS TECHNOLOGY IS BASED ON THE DISCOVERY THAT THE 5' NUCLEASE OF SEVERAL THERMOSTABLE DNA POLYMERASES CAN BE ENGINEERED TO DIRECT CLEAVAGE OF DNA OR RNA AT ONE AND ONLY ONE LOCATION, ALLOWING THE DEVELOPMENT OF A VERY FAST, SENSITIVE, SPECIFIC, AND ISOTHERMAL CLEAVAGE-BASED DETECTION TECHNOLOGY. PHASE I SHOULD SHOW THAT THE SIGNAL RELEASED FROM THE RECOGNITION OF THE TARGET SEQUENCE CAN BE PROPAGATED THROUGH AN AMPLIFYING CLEAVAGE CASCADE. THIS WILL ENTAIL OPTIMIZATION OF CONDITIONS FOR CLEAVAGE, FURTHER CHARACTERIZATION OF THE ENZYME ACTIVITY, AND TESTING OF THE MOLECULES INVOLVED IN THE AMPLIFICATION CASCADE. PHASE II WOULD COMPLETE THE DEVELOPMENT OF THIS BROAD-BASED NUCLEIC ACID DETECTION TECHNOLOGY THAT CAN PLAY A SIGNIFICANT ROLE IN MAKING COST-EFFECTIVE, LARGE-SCALE EARLY DETECTION, DIAGNOSIS, AND MONITORING OF A WIDE VARIETY OF DISEASES A REALITY.