Genome editing in bread wheat toward reducing allergenic gluten

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K05977
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 39010:Science in plant genetics and breeding-related
• Research Institution: Yokohama City University
• Principal Investigator: Kawaura Kanako 横浜市立大学, 木原生物学研究所, 准教授 (60381935)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: パンコムギ / ゲノム編集 / グルテン / 転写因子 / 小麦アレルギー

Outline of Final Research Achievements

In wheat, gliadins are seed storage proteins and major components of gluten in flour. The copy number of genes encoding gliadins is particularly high, and some of these proteins are allergenic. We previously reported that the expression of the multiple genes is independently regulated in seed developmental stage. In this study, CRISPR/Cas9 genome editing technique was applied to identify transcription factors that regulate the expression of each gliadin gene. In addition, genome editing of candidate transcription factors, namely SPA and SHP, was performed to establish hypoallergenic wheat lines. Since we could not obtain mutant lines by introducing Cas9 and gRNA RNPs, we conducted the experiment by introducing plasmid vectors into immature embryos using the particle delivery system. As a result, lines with mutations in three homoeologous genes were obtained in SPA, however no mutant line was obtained in SHP. Conditions for increasing genome editing efficiency need to be determined.

Free Research Field

植物ゲノム科学

Academic Significance and Societal Importance of the Research Achievements

コムギは重要作物であるが、アレルギーや自己免疫疾患の原因として問題となる。コムギでもゲノム編集技術が使えるようになったが、アレルギーの原因となるグルテンタンパク質をコードする遺伝子はゲノム中で高度に重複して存在するため、個別にゲノム編集することは現実的でない。本研究では、転写因子のゲノム編集により包括的に多重遺伝子の発現制御を行うことで、低アレルゲン小麦系統作出に向けた方略を提案できると考える。