** AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** NOROVIRUS (NOV) IS AN EXTREMELY PREVALENT FOODBORNE PATHOGEN THAT CAUSES SIGNIFICANT HUMAN AND ECONOMIC LOSSES EVERY YEAR. CURRENT DETECTION METHODS, RELYING ON POLYMERASE CHAIN REACTION (PCR) TECHNOLOGY, ARE COMPLEX AND OFTEN INACCESSIBLE. THIS PROJECT PROPOSES A NOVEL SOLUTION: A CRISPR-CAS14 G-QUADRUPLEX (CRISPR-G4) BIOSENSOR FOR ON-SITE DETECTION OF NOV IN LEAFY GREENS. OUR SYSTEM FIRST AMPLIFIES NOV RNA ISOTHERMALLY USING LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP). WE WILL THEN UTILIZE A CRISPR-CAS14 SYSTEM FOR EXTREMELY SPECIFIC AND ROBUST TARGET RECOGNITION. THIS SYSTEM PROVIDES A FRONTIER INTO THE NEXT GENERATION OF CRISPRBASED BIOSENSORS, DRAWING ON THE COMPACT SIZE AND BIOSENSING CAPABILITIES OF THE CAS14 ENZYME. THE SYSTEM IS THEN COMBINED WITH G4S TO PRODUCE A SIMPLE COLORIMETRIC READOUT. THIS APPROACH REQUIRES NO SPECIALIZED EQUIPMENT, ENABLING RAPID AND AFFORDABLE ON-SITE TESTING. WE PROPOSE THREE OBJECTIVES, INCLUDING:1) THE DESIGN OF THE CRISPR-CAS14 SYSTEM FOR THE DETECTION OF NOV RNA,2) THE DEVELOPMENT OF THE CRISPR-G4 SYSTEM,3) VALIDATION OF THE EFFICACY IN DETECTING NOV IN LEAFY GREENS.BASED ON OUR EXPERTISE AND PREVIOUS LITERATURE, WE ANTICIPATE THAT THE PROPOSED BIOSENSOR WILL PROVIDE EXTREMELY SENSITIVE AND SPECIFIC RECOGNITION OF NOV IN SPINACH AND LETTUCE AND COULD POTENTIALLY BE EXTENDED FOR SIMILAR SYSTEMS. THE PROJECT ALIGNS SEAMLESSLY WITH THE FOOD SAFETY NUTRITION, AND HEALTH PRIORITY AREA DUE TO THE POTENTIAL OF OUR PROPOSED SENSOR TO COMBAT FOODBORNE ILLNESSES. WE ANTICIPATE THAT OUR SENSOR WILL SUBSTANTIALLY ADDRESS THE URGENT NEED FOR ACCESSIBLE NOV TESTING, POTENTIALLY REDUCING FOODBORNE ILLNESSES AND ECONOMIC LOSS.