DEVELOPMENT OF CRISPR-EQUIPPED ENGINEERED PHAGES (CREEPS) FOR THE DETECTION OF ESCHERICHIA COLI (INDICATOR BACTERIA) IN AGRICULTURAL WATER

In this project, we aim to develop a state-of-the-art CRISPR-equipped engineered phage (CREEP) for the rapid and sensitive detection ofEscherichia coli(E. coli) that serves as microbial contamination indicators in agricultural water. The cutting-edge detection assay will provide fresh produce workers with an easy-to-use tool to estimate the microbial safety of both irrigation and washing waters that come in direct contact with fresh produce. This is important for the long-term goal of preventing the spread of foodborne pathogens in the fresh produce supply chains and reducing foodborne outbreaks. The specific objectives include 1) design and optimize the CRISPR-Cas12a system to detect bacterial genomic DNA (gDNA), 2) fabricate CRISPR-equipped engineered phages (CREEP) to detect E. coli, and 3) validate CREEPs to estimate the microbial safety in agricultural water.