期刊
PLANT BIOTECHNOLOGY JOURNAL
卷 16, 期 4, 页码 902-910出版社
WILEY
DOI: 10.1111/pbi.12837
关键词
coeliac disease; alpha-gliadins; CRISPR/Cas9
资金
- Spanish Ministry of Economy and Competitiveness [AGL2016-80566-P, AGL2013-48946-C3-1-R]
- Fundacion Alfonso Martin Escudero
- European Regional Development Fund (FEDER)
Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The alpha-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 alpha-gliadin genes. Twenty-one mutant lines were generated, all showing strong reduction in alpha-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.
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