Wheat is one of the most widely cultivated crops and serves as a staple food for millions of people. A key component is gluten—a variable protein complex including α/β-, γ-, and ω-gliadins. While gluten is harmless for most, it can cause health issues in individuals with Celiac Disease (CD), Non-Celiac Wheat Sensitivity (NCWS), and IgE-mediated food allergies which may require gluten-free foods in their diet.

Francisco Barro, Institute for Sustainable Agriculture (IAS-CSIC), Córdoba, Spain, and colleagues have used CRISPR-Cas9 technology to create wheat with significantly reduced gluten content, advancing the production of gluten-free wheat.

By applying CRISPR/Cas multiplexing, the team introduced targeted mutations into γ- and ω-gliadin genes of wheat, to produce lines deficient in one or both immunogenic gliadin fractions simultaneously.

They designed eight single guide RNAs (sgRNAs) and combined them into four plasmids to produce 59 modified wheat lines, of which 20 showed mutations in the target genes. Characterization of these lines through Sanger or NGS sequencing revealed a complex pattern of InDels, including deletions spanning multiple sgRNAs.

The mutations were inherited by the offspring and the gluten content was reduced by up to 97.7%. When these lines were crossed with other CRISPR-edited lines with reduced α-gliadins, these lines produced very low to no gluten content.

The researchers say that their work represents an important step towards producing gluten-free wheat, and they plan to conduct further studies to assess how these modifications affect the immune response in CD and NCWS patients.

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• CRISPR/Cas9-mediated multiplex gene editing of gamma and omega gliadins, paving the way for gliadin-free wheat,
  Susana Sánchez-León, Miriam Marín-Sanz, María H. Guzmán-López, Marta Gavilán-Camacho, Edurne Simón, Francisco Barro,
  Journal of Experimental Botany 2024, erae376.
  https://doi.org/10.1093/jxb/erae376