** AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** THE PROPOSED PROJECT ADDRESSES THE PROGRAM AREA PLANT BREEDING FOR AGRICULTURAL PRODUCTION (A1141). THE LONG-TERM GOAL OF THIS RESEARCH IS TO REMOVE IMMUNOGENIC-GLUTEN PROTEINS, NOT WHEAT, FROM THE DIET. THIS GOAL WILL BE ACHIEVED BY DEVELOPING REDUCED IMMUNOGENICITY, LYSINE-RICH WHEAT GENOTYPES USING CONVENTIONAL AND GENOME-EDITING METHODS. THESE REDUCED-IMMUNOGENICITY GENOTYPES OFFER AN EASY-TO-INTEGRATE SOLUTION IN THE CURRENT PRODUCTION, PROCESSING, AND DISTRIBUTION SYSTEMS, HENCE OFFERING AN AFFORDABLE SOLUTION TO ALL IRRESPECTIVE OF THEIR SOCIOECONOMIC STRATA. THE FOLLOWING SPECIFIC OBJECTIVES WILL BE UNDERTAKEN TO ACHIEVE THE PROPOSED GOAL. 1) DEVELOP NON-IMMUNOGENIC WHEAT GENOTYPES BY STACKING NULL-MUTANTS AT WHEAT GLIADIN AND GLUTENIN LOCI. THIS OBJECTIVE WILL BE ACHIEVED BY THE GENETIC CROSSING OF REDUCED-GLUTEN MUTANTS OR USING A NOVEL COMBINATION OF MULTI-GENE EDITING AND NANOPARTICLE-BASED GENE DELIVERY METHOD. 2) EVALUATE SELECTED REDUCED-IMMUNOGENICITY WHEAT GENOTYPES FOR SUITABILITY FOR HUMAN CONSUMPTION. THIS OBJECTIVE WILL BE ACHIEVED BY A DETAILED CHARACTERIZATION OF WHEAT GENOTYPES WITH STACKED GLIADIN AND GLUTENIN NULL MUTANTS FOR THEIR BIOCHEMICAL AND IMMUNOLOGICAL PROPERTIES USING ELECTROPHORESIS, LIQUID CHROMATOGRAPHY, MASS SPECTROMETRY, AND IMMUNOLOGICAL ASSAYS (ELISA AND T-CELL). SELECTED LINES WILL BE ACCESSED FOR END-USE PERFORMANCE. 3) ASSESS THE SOCIAL AND COMMERCIAL ACCEPTANCE OF THE REDUCED-IMMUNOGENICITY WHEAT LINES. THE OBJECTIVE WILL BE ACHIEVED BY SURVEYING CONSUMERS' OPINIONS, INCLUDING CELIAC PATIENTS, WHEAT PRODUCERS, MILLERS, AND BAKERS, ON USING GENOME EDITING TO DEVELOP REDUCED-IMMUNOGENICITY, HIGH-LYSINE WHEAT LINES. THESE INTERACTIONS WILL ALLOW RESEARCHERS TO ENHANCE THEIR UNDERSTANDING OF THE PUBLIC PERCEPTION OF THE PROPOSED SOLUTIONS AND CONSIDER THIS INFORMATION TO READJUST THE RESEARCH BASED ON PUBLIC OPINION.