CRISPR/CAS9-MEDIATED GENOME EDITION (GE) IN FOOD-ANIMALS CAN BENEFIT BOTH THE SCIENTIFIC COMMUNITY AND THE FUTURE AGRICULTURAL INDUSTRY. THE CURRENT METHOD FOR GE IN CHICKENS REQUIRES HIGHLY SKILLED EXPERTISE FOR: 1) ISOLATION, CULTURE, AND GE OF PRIMORDIAL GERM CELLS (PGCS) IN VITRO, 2) TRANSFER OF GENOME-EDITED PGCS INTO RECIPIENT EMBRYOS, AND 3) GENERATION OF GERMLINE CHIMERAS AND ITS OFFSPRING WITH GE. IN ADDITION, THE CURRENT PGC-MEDIATED METHOD HAS NOT BEEN SUCCESSFUL FOR OTHER AVIAN SPECIES. THEREFORE, IT IS NECESSARY TO DEVELOP AN EASY AND EFFICIENT METHOD WITHOUT MEDIATING PGCS IN CULTURE. TO IMPROVE EFFICIENCY AND OVERCOME THESE TECHNICAL BARRIERS, WE DEVELOPED AN OPTIMIZED CRISPR/CAS9 VECTOR AND GENERATED GENOME-EDITED POULTRY USING ADENOVIRUS AS A DELIVERY SYSTEM OF THE OPTIMIZED CRISPR/CAS9 INTO BLASTODERMS WITHOUT MEDIATING PGC IN CULTURE. THE EFFICIENCY OF TARGETED GENOME MODIFICATION IN POULTRY DEPENDS ON THE RATE OF GENOME MODIFICATION IN THE GERM CELL POPULATION. THEREFORE, THIS PROPOSAL IS FOCUSING ON THE DEVELOPMENT OF A STRATEGY TO DIRECTLY TARGET MORE GERM CELLS BY INJECTING THE CRISPR/CAS9 ADENOVIRUS INTO THE EMBRYO. WITH FOUR OBJECTIVES, WE WILL INJECT THE CRISPR/CAS9 ADENOVIRUS INTO FOUR DIFFERENT LOCATIONS WHERE PGCS ARE DISPERSED, CIRCULATING OR CONGREGATED DURING EARLY EMBRYO DEVELOPMENTAL PROCESSES TO PROVIDE THE MOST EFFICIENT NEXT-GENERATION METHOD FOR GE IN POULTRY. SUCCESSFUL DEVELOPMENT OF THE PROPOSED NEXT GENERATION METHOD OF GE IN POULTRY IS EXPECTED TO ADVANCE AVIAN RESEARCH AND DEVELOP MORE PRODUCTIVE GENOMIC LINES OF POULTRY FOR INFECTIOUS DISEASE RESISTANCE, HIGHER GROWTH AND FEED EFFICIENCY, AND INCREASED MUSCLE WITH REDUCED FAT.