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CRISPR-based editing of the omega- and gamma-gliadin gene clusters reduces wheat immunoreactivity without affecting grain protein quality

posted on 2024-06-11, 05:08 authored by Kansas State University
Wheat immunotoxicity is associated with abnormal reaction to gluten-derived peptides. Attempts to reduce immunotoxicity using breeding and biotechnology often affect dough quality. Here, the multiplexed CRISPR-Cas9 editing of cultivar Fielder was used to modify gluten-encoding genes, specifically focusing on omega- and gamma-gliadin gene copies, which were identified to be abundant in immunoreactive peptides based on the analysis of wheat genomes assembled using the long-read sequencing technologies. The whole genome sequencing of an edited line showed mutation or deletion of nearly all omega-gliadin and half of the gamma-gliadin gene copies and confirmed the lack of editing in the alpha/beta-gliadin genes. The estimated 75% and 64% reduction in omega- and gamma-gliadin content, respectively, had no negative impact on the end-use quality characteristics of grain protein and dough. A 47-fold immunoreactivity reduction compared to non-edited line was demonstrated using antibodies against immunotoxic peptides. Our results indicate that the targeted CRISPR-based modification of the omega- and gamma-gliadin gene copies determined to be abundant in immunoreactive peptides by analyzing high-quality genome assemblies is an effective mean for reducing immunotoxicity of wheat cultivars while minimizing the impact of editing on protein quality.


National Institute of Food and Agriculture, 2020-67013-30906

National Institute of Food and Agriculture, 2021-67013-34174

Bill and Melinda Gates Foundation, INV-004430


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BioProject Curation Staff


National Center for Biotechnology Information

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  • Non-geospatial

ISO Topic Category

  • biota

National Agricultural Library Thesaurus terms

sequence analysis

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  • No

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  • Public

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It is recommended to cite the accession numbers that are assigned to data submissions, e.g. the GenBank, WGS or SRA accession numbers. If individual BioProjects need to be referenced, state that "The data have been deposited with links to BioProject accession number PRJNA1044136 in the NCBI BioProject database ("

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