Departamento de Mejora Genética Vegetal, Instituto de Agricultura Sostenible (IAS-CSIC), Córdoba, Spain.
Department of Genetics, Cell Biology, and Development, Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
Plant Biotechnol J. 2018 Apr;16(4):902-910. doi: 10.1111/pbi.12837. Epub 2017 Nov 24.
Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The α-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 α-gliadin genes. Twenty-one mutant lines were generated, all showing strong reduction in α-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.
乳糜泻是一种自身免疫性疾病，在遗传易感性个体中，由摄入小麦、大麦和黑麦中的麸质蛋白触发。小麦的α-麦醇溶蛋白基因家族包含四个高度刺激的肽，其中 33 肽是乳糜泻患者的主要免疫显性肽。我们设计了两个 sgRNA 来靶向α-麦醇溶蛋白基因中编码序列附近的保守区域。生成了 21 条突变系，所有系的α-麦醇溶蛋白均明显减少。在鉴定出的 45 个野生型基因中，其中一条突变系的突变多达 35 个不同的基因，而免疫反应性降低了 85%。鉴定出无转基因的系，并且在任何潜在的靶标中都没有检测到脱靶突变。这里描述的低麸质、无转基因的小麦系可用于生产低麸质食品，并作为材料将该特性导入优良小麦品种。
