Development of Simple Detection Method of the Gene Edited Low Gluten Wheat by Homo/Hetero-Duplexes and T7E1 Technique
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Abstract
The development and application of agricultural gene-editing technology have been studied in various crops. The current regulation framework in Thailand is still under consideration. Therefore, the ability to detect gene-edited plants is essential for monitoring and verifying such plants as part of the preparation for the adoption of this technology. This study focuses on the research and development of an investigative method for detecting gene-edited wheat in a simpler and faster manner. Wheat with edited α-gliadin family genes in the sgAlpha-2 region, a group of genes responsible for controlling wheat gluten production, was used in this study. The gene editing involved the deletion of only a few bases from the genome, resulting in a significant reduction in gluten protein levels in the edited wheat. The homo/hetero duplex and T7 endonuclease-I (T7E1) assay was employed as the detection technique. This method relies on the principle of pairing complementary and non-complementary DNA strands. Mismatched DNA pairs form a characteristic loop in the editable region, which can be detected using gel electrophoresis analysis to differentiate DNA bands. This approach enables the identification of gene-edited plant samples effectively.
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References
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