THE WESTERN FLOWER THRIPS, FRANKLINIELLA OCCIDENTALIS, IS GLOBAL PEST AND A VECTOR OF TOMATO SPOTTED WILT VIRUS (TSWV) AND RELATED ORTHOTOSPOVIRUSES THAT CAUSE SIGNIFICANT YIELD LOSSES IN CROPS. OUR LONG-RANGE GOAL IS TO DEVELOP GENOME-EDITING TECHNOLOGIES FOR POPULATION SUPPRESSION AND/OR POPULATION REPLACEMENT OF F. OCCIDENTALIS AS A MEANS OF REDUCING RELIANCE ON CONVENTIONAL CHEMICAL MANAGEMENT STRATEGIES AND MITIGATING ORTHOTOSPOVIRUS TRANSMISSION BY THIS SUPERVECTOR. GENOME-EDITING HAS NOT YET BEEN APPLIED TO THRIPS AND APPLICATION OF THESE TECHNOLOGIES WILL ALSO ENABLE BASIC STUDIES OF GENE FUNCTION IN THRIPS, INCLUDING VIRUS-VECTOR INTERACTIONS. THE AIM OF THIS PROPOSAL IS TO DEVELOP AND OPTIMIZE A PROOF-OF-PRINCIPLE CRISPR/CAS9 GENE-KNOCKOUT SYSTEM FOR F. OCCIDENTALIS USING TRADITIONAL AND NEW METHODOLOGIES OF DELIVERY OF CAS9 AND GUIDE RNAS. WE WILL TARGET GENES WITH EASILY SCORED VISIBLE PHENOTYPES WHICH WILL ENABLE DIRECT FEEDBACK THEREBY HASTENING PROGRESS AND CLEARLY SIGNALING SUCCESS. THE SPECIFIC RESEARCH OBJECTIVES ARE TO:OBJECTIVE 1. DEVELOP METHODS FOR MICROINJECTING PRECELLULAR F. OCCIDENTALIS EMBRYOS.OBJECTIVE 2. USE THE CRISPR/CAS9 SYSTEM TO KNOCKOUT F. OCCIDENTALIS EYE-COLOR GENES.OUR RESEARCH ADDRESSES THE USDA-NIFA CHALLENGE OF DEVELOPING MORE SUSTAINABLE, PRODUCTIVE AND ECONOMICALLY-VIABLE PLANT PRODUCTION SYSTEMS. SPECIFICALLY, WE WILL EXPAND THE USE OF GENE EDITING TECHNOLOGY TO VECTORS OF PLANT VIRUSES FOR PEST AND DISEASE CONTROL.