Engineered dsRNA–protein nanoparticles for effective systemic gene silencing in plants
Huayu Sun1,9,† , Ankarao Kalluri2,† , Dan Tang1 , Jingwen Ding3 , Longmei Zhai1 , Xianbin Gu1 , Yanjun Li1 , Huseyin Yer1 , Xiaohan Yang4 , Gerald A. Tuskan4 , Zhanao Deng5 , Frederick G. Gmitter Jr6 , Hui Duan7 and Challa Kumar2,3,8, , Yi Li,1,
1Department of Plant Science and Landscape Architecture, University of Connecticut, Storrs, CT 06269, USA 2Department of Material Science, University of Connecticut, Storrs, CT 06269, USA 3Department of Chemistry, University of Connecticut, Storrs, CT 06269, USA 4Biosciences Division, Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA 5Gulf Coast Research and Education Center, University of Florida, IFAS, Wimauma, FL 33598, USA 6Citrus Research and Education Center, University of Florida, IFAS, Lake Alfred, FL 33850, USA 7USDA-ARS, U.S. National Arboretum, Floral and Nursery Plants Research Unit, Beltsville Agricultural Research Center (BARC)-West, Beltsville, MD 20705, USA 8Department of Molecular and Cellular Biology, University of Connecticut, Storrs, CT 06269, USA 9Institute of Gene Science and Industrialization for Bamboo and Rattan Resources, International Center for Bamboo and Rattan, Beijing 100102, China *Corresponding author. E-mail: challa.kumar@uconn.edu,yi.li@uconn.edu †Both authors contributed equally to the study.
Received: 27 Dec 2023 Accepted: 06 Feb 2024 Published online: 22 Feb 2024
Abstract
Long-distance transport or systemic silencing effects of exogenous biologically active RNA molecules in higher plants have not been reported. Here, we report that cationized bovine serum albumin (cBSA) avidly binds double-stranded beta-glucuronidase RNA (dsGUS RNA) to form nucleic acid–protein nanocomplexes. In our experiments with tobacco and poplar plants, we have successfully demonstrated systemic gene silencing effects of cBSA/dsGUS RNA nanocomplexes when we locally applied the nanocomplexes from the basal ends of leaf petioles or shoots. We have further demonstrated that the cBSA/dsGUS RNA nanocomplexes are highly effective in silencing both the conditionally inducible DR5-GUS gene and the constitutively active 35S-GUS gene in leaf, shoot, and shoot meristem tissues. This cBSA/dsRNA delivery technology may provide a convenient, fast, and inexpensive tool for characterizing gene functions in plants and potentially for in planta gene editing.
, Yi Li,1 ,
