THIS PROJECT UTILIZES A NON-INFECTIOUS PLANT VIRUS NANOPARTICLE FORDELIVERY OF MRNA INTO PLANTS, ALLOWING FOR A TEMPORARY CHANGE TO THE BEHAVIOR OF THE PLANTS WITHOUT PERMANENTLY ALTERING THEIR GENETIC MATERIAL. THIS CAN ALLOW PLANTS TO BE TEMPORARILY EQUIPPED TO RESIST DISEASE, OVERCOME DROUGHT CONDITIONS, OR PERFORM A NEW FUNCTION WITHOUT CHANGING ITS DNA OR CREATING REGULATORY HURDLES FOR PRODUCTION. AS THE HUMAN POPULATION CONTINUES TO GROW, IT IS IMPORTANT THAT WE HAVE MORE TECHNIQUES TO ADDRESS FOOD SECURITY, AND MITIGATING THE LOSS OF CROPS IS ONE OF THE MOST CHALLENGING ISSUES IN THIS AREA. OUR ULTIMATE GOAL IS TO STIMULATE A COMMON IMMUNE RESPONSE FOUND IN A MANY PLANTS USING THIS TECHNIQUE SO WE CAN HAVE A GENERALIZED PLANT VACCINE. IN MANY WAYS, THE PRINCIPLES OF THIS TECHNOLOGY ARE SIMILAR TO THOSE UTILIZED IN THE LIPID NANOPARTICLE COVID-19 VACCINES, BUT ADAPTED FOR USE IN PLANTS. IN THIS WORK, WE WILL FIRST WORK TO ENGINEER THE VIRUS NANOPARTICLES TO DELIVER THE APPROPRIATE MRNA BY BREAKING THEM APART, REMOVING THEIR ORIGINAL GENETIC MATERIAL, AND REPLACING IT WITH MRNA FOR THE EXPRESSION OF A GREEN FLUORESCENT PROTEIN BEFORE REASSEMBLING THE PARTICLES. WE WILL THEN INTRODUCE THESE PARTICLES TO A CULTURE OF CELLS DERIVED FROM PLANTS AS A PROOF OF CONCEPT. THE CELLS THAT UPTAKE THE VIRUSES SHOULD BEGIN TO GLOW GREEN AS THEY EXPRESS THE DELIVERED MRNA, AND WE WILL USE THIS INFORMATION TO SEE HOW EFFICIENT THE DELIVERY IS AND HOW LONG THE CELLS STAY GREEN. AFTER SOME OPTIMIZATION, WE WILL APPLY THESE NANOPARTICLES DIRECTLY TO SEVERAL PLANTS USING ABRASION, INJECTION, AND PASSIVE DIFFUSION TO SEE IF THEIR TISSUES BEGIN TO GLOW GREEN AS WELL. WITH THIS INFORMATION, WE CAN SEE IF ANY OF THE NANOPARTICLES NEED TO BE IMPROVED FOR HIGHER DELIVERY EFFICIENCY. IF THEY DO NEED TO BE MODIFIED, WE WILL ADD CELL-PENETRATING PEPTIDES TO THEIR SURFACE WHICH ALLOW THE NANOPARTICLES TO EMBED THEMSELVES IN THE CELL WALL AND MEMBRANE OF THE PLANTS FOR FASTER UPTAKE. ONCE WE HAVE SUCCESSFULLY DEMONSTRATED OUR TECHNOLOGY USING THE GREEN FLUORESCENT PROTEIN, WE WILL MOVE ON TO DELIVERING SOME TRANSCRIPTS IN OUR NANOPARTICLES THAT MAY STIMULATE AN IMMUNE RESPONSE IN PLANTS. WE WILL USE ANALYTICAL TECHNIQUES TO SEE IF THE EXPRESSION OF DIFFERENT GENES, THE METABOLISM, AND THE PROTEINS IN THESE PLANT CELLS HAVE CHANGED DUE TO THE DELIVERY OF THESE TRANSCRIPTS. IF THE RESULTS INDICATE WE MAY HAVE ACTIVATED AN IMMUNE RESPONSE IN CELL CULTURE, WE WILL ADVANCE TO WHOLE PLANT DELIVERY AND USE THE SAME ANALYTICAL TECHNIQUES, IMAGING, AND HISTOLOGY TO DETERMINE THE EFFICIENCY OF OUR TECHNOLOGY. IT IS OUR HOPE THAT THIS TECHNOLOGY WILL ENABLE BROAD PROTECTION AGAINST MANY DIFFERENT PATHOGENS USING THE HIGHLY CONSERVED SYSTEMIC ACQUIRED RESISTANCE PATHWAY FOUND IN A LARGE NUMBER OF PLANTS. SPECIFICALLY, WE HOPE THIS CAN BE USED AS AN ON-DEMAND VACCINE FOR PLANTS WHEN INFECTION IS DETECTED SOMEWHERE IN THE FIELD TO PROTECT THE REST OF THE CROPS, WHICH WOULD SIMULTANEOUSLY PREVENT CROP LOSSES AND IMPROVE THE ECONOMIC OUTLOOK OF FARMERS IN DISEASE-PRONE AREAS.