D3416: Development of targeted phage-based antimicrobials to maximize the production and health of Anastrepha ludens

Anastrepha ludens is a key pest of citrus and other fruit crops. Large scale rearing is currently employed for sterile insect release to improve containment at the Texas-Mexico border. The synthetic feeding systems utilized in insect rearing represent an ideal breeding ground for bacterial colonization and growth. Among bacteria identified within these artificial diets are human and insect pathogens, as well as beneficial bacteria that improve the production rate, health, and mating success of insects. Here we investigate the prophylactic use of bacteriophage-based therapies for the control of these pathogens and the retention of beneficial microbial flora. We have designed this system based on a recently discovered adaptive immune system found among many Bacteria and Archaea species called CRISPR-Cas. While these systems have gained widespread attention as powerful genome-editing tools in eukaryotic cells, we showed that they can be co-opted as programmable and potent antimicrobials. Our system, in effect, tricks the CRISPR-Cas system into recognizing the pathogen genome as a foreign invader, resulting in species-specific killing of the pathogen. The development of targeted antimicrobials for known entomopathogens can reduce the loss, and thus the cost, of mass rearing insects. Moreover, because our proposed antimicrobials will be targeted for individual pathogens they can be used prophylactically without risking the health of the insect due to loss of beneficial bacteria. We believe that this approach can produce a healthier, more attractive, sterile male insect for less money than current methods.