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ARS Home » Plains Area » Houston, Texas » Children's Nutrition Research Center » Research » Publications at this Location » Publication #375039

Research Project: Nutritional Role of Phytochemicals

Location: Children's Nutrition Research Center

Title: Expression of small interfering RNAs in lettuce using artificial microRNA technology

item KAKESHPOUR, TAYEBEH - Kansas State University
item TAMANG, TEJ MAN - Kansas State University
item PARK, WOOJUN - Baylor College Of Medicine
item MANOHAR, MURLI - Cornell University
item YANG, JIAN - Children'S Nutrition Research Center (CNRC)
item HIRSCHI, KENDAL - Children'S Nutrition Research Center (CNRC)
item PARK, SUNGHUN - Kansas State University

Submitted to: Biotechniques
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/3/2020
Publication Date: 1/5/2020
Citation: Kakeshpour, T., Tamang, T., Park, W.D., Manohar, M., Yang, J., Hirschi, K.D., Park, S. 2020. Expression of small interfering RNAs in lettuce using artificial microRNA technology. Biotechniques. 68(4).

Interpretive Summary: This study is working towards developing plant-based diets for gene therapy. Some vaccines attempt to delivery small RNAs to thwart viruses like COVID-19. In the future, we want to address if diets can be used to deliver therapeutic small RNAs. The knowledge gap we are addressing here is how to make large amounts of artificial small RNAs in plants. In this work, we developed a series of expression techniques to produce large amounts of foreign small RNAs in the edible portions of lettuce. These foreign RNAs were shown to be stable in the engineered lettuce lines. This work demonstrates the feasibility of having plants produce large amounts to therapeutic small RNAs.

Technical Abstract: Artificial miRNA technology enables the generation of siRNAs to regulate the expression of targeted genes. However, the application of siRNAs to alter gene expression is challenging due to their instability and requires a means to efficiently deliver siRNAs into the host. Here, we report that the siRNAs targeted to animal mRNAs can be heterologously expressed and stably produced in lettuce. We have modified rice miRNA precursors to produce siRNAs in lettuce with the potential to target mRNAs of mouse complement 3 (C3) and coagulation factor 7 (CF7). Expression of primary and mature siRNAs in the transgenic lettuce lines was confirmed via Sanger sequencing. Our study demonstrates an applicable tool to alter gene expression in the targeted host and has potential utility in siRNA-based oral therapeutics.