DEVELOPING WHEAT GENOTYPES WITH REDUCED IMMUNOGENICITY AND ENHANCED LYSINE CONTENT

The long-termgoal 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 gluten-free grains and products production, processing, and distribution systems; hence, they will offer an affordable solution to all, irrespective of their socioeconomic strata. Further, the reduced immunogenicity and lysine-rich wheat genotypes will benefit both gluten-sensitive and healthy individuals equally, as eliminating gliadins and LMW-GS reduce immunogenicity and simultaneously enhance grain lysine content and improve digestibility and bioavailability. 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. Theworking hypothesisis that we will gain the desired reduction in the gluten content of wheat lines via the pyramiding of prolamin null mutants.2)Evaluate selected reduced-immunogenicity wheat genotypes for suitability for human consumption.This objective could 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. Theworking hypothesisis that the immunological assays will provide strong evidence for the safety of non-immunogenic wheat lines for celiac patients.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 feedback this information to readjust the research based on public opinion (i.e., acceptance level) and inform the general public about the pros and cons of the current therapy and compare it with the proposed solution. Theworking hypothesisis that public engagement is vital for the successful implementation of a treatment.