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Sequencing of genome-edited wheat lines derived from cultivar Fielder

dataset
posted on 2024-01-27, 04:50 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.

Funding

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

History

Data contact name

BioProject Curation Staff

Publisher

National Center for Biotechnology Information

Temporal Extent Start Date

2023-11-12

Theme

  • Non-geospatial

ISO Topic Category

  • biota

National Agricultural Library Thesaurus terms

sequence analysis

Pending citation

  • No

Public Access Level

  • Public

Accession Number

PRJNA1039537

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