Gluten is composed of glutenins and gliadins and determines the viscoelastic properties of dough and end-use quality in wheat (Triticum aestivum L.). Gliadins are important for wheat end-use traits, but the contribution of individual gliadin genes is unclear, since gliadins are encoded by a complex, multigenic family, including many pseudogenes.We used CRISPR/Cas9-mediated gene editing and map-based cloning to investigate the contribution of the c-gliadin genes annotated in the wheat cultivar 'Fielder', showing that Gli-c1-1D and Gli-c2-1B account for most of the c-gliadin accumulation.The impaired activity of only two c-gliadin genes in knockout mutants improved end-use quality and reduced gluten epitopes associated with celiac disease (CD). Furthermore, we identified an elite haplotype of Gli-c1-1D linked to higher end-use quality in a wheat germplasm collection and developed a molecular marker for this allele for marker-assisted selection.Our findings provide information and tools for biotechnology-based and classical breeding programs aimed at improving wheat end-use quality.