Low-gluten, nontransgenic wheat engineered with CRISPR/Cas9.

ABSTRACT: Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The ?-gliadin gene family of wheat contains four highly stimulatory peptides, of which the 33-mer is the main immunodominant peptide in patients with coeliac. We designed two sgRNAs to target a conserved region adjacent to the coding sequence for the 33-mer in the ?-gliadin genes. Twenty-one mutant lines were generated, all showing strong reduction in ?-gliadins. Up to 35 different genes were mutated in one of the lines of the 45 different genes identified in the wild type, while immunoreactivity was reduced by 85%. Transgene-free lines were identified, and no off-target mutations have been detected in any of the potential targets. The low-gluten, transgene-free wheat lines described here could be used to produce low-gluten foodstuff and serve as source material to introgress this trait into elite wheat varieties.

SUBMITTER: Sanchez-Leon S 

PROVIDER: S-EPMC5867031 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature