DEVELOPING AN INFRASTRUCTURE AND PRODUCT TEST PIPELINE TO DELIVER NOVEL THERAPIES FOR CITRUS GREENING DISEASE

1. Protein-protein interaction blockers-peptides and RNA aptamers We propose to couple co-immunoprecipitation (coIP) and targeted proteomics to identify the receptors within the psyllid that interact with the gut binding peptides characterized by the Shatters Lab. The peptides used in this work were hexameric peptides with C-terminus ending in Gly-Gly-biotin linker-ligand. Using quantitative mass spectrometry, differences in protein stoichiometry can be determined in addition to the reliable identification of specific binding partners. Next, we will use a targeted proteomic platform, which includes selected reaction monitoring (SRM) and co-IP to specifically quantify the level of the gut surface receptors during CLas infection of the psyllid, in different psyllid tissues, during psyllid development, in different psyllid populations, and during RNAi experiments to silence the receptors. 2. RNAi- Oral delivery of dsRNAs as RNAi inducers. Drs. Shatters, Hunter and Stover conduct research on the use of orally delivered dsRNAs as RNAi-inducers that produce phenotypes in the psyllid that could be adapted as psyllid control strategies (mortality, development malformations, blocking CLas transmission, etc. This project will focus only on topical application strategies for therapeutic delivery of dsRNA exogenously applied to citrus and subsequent systemic movement of the dsRNA in the plant followed by psyllid uptake during feeding. Research will be conducted to identify the best targets or combination of targets and most effective candidates will be tested in field trees using several application strategies that will be optimized to support dsRNA stability and efficient dsRNA uptake by the plant. 3. Delivery Strategies Therapeutic Molecule Delivery. Drs. Shatters and Stover have obtained successes in optimizing therapeutic delivery of â¿¿antibioticâ¿Â