Engineered dsRNA–protein nanoparticles for effective systemic gene silencing in plants

Author:

Sun Huayu12ORCID,Kalluri Ankarao3,Tang Dan1,Ding Jingwen4,Zhai Longmei1,Gu Xianbin1,Li Yanjun1,Yer Huseyin1,Yang Xiaohan5,Tuskan Gerald A5,Deng Zhanao6,Gmitter Jr Frederick G7,Duan Hui8ORCID,Kumar Challa342,Li Yi1

Affiliation:

1. University of Connecticut Department of Plant Science and Landscape Architecture, , Storrs, CT 06269, USA

2. International Center for Bamboo and Rattan Institute of Gene Science and Industrialization for Bamboo and Rattan Resources, , Beijing 100102, China

3. University of Connecticut Department of Material Science, , Storrs, CT 06269, USA

4. University of Connecticut Department of Chemistry, , Storrs, CT 06269, USA

5. Oak Ridge National Laboratory Biosciences Division, Center for Bioenergy Innovation, , Oak Ridge, TN 37831, USA

6. University of Florida, IFAS Citrus Research and Education Center, , Lake Alfred, FL 33850, USA

7. Beltsville Agricultural Research Center (BARC)-West USDA-ARS, U.S. National Arboretum, Floral and Nursery Plants Research Unit, , Beltsville, MD 20705, USA

8. University of Connecticut Department of Molecular and Cellular Biology, , Storrs, CT 06269, USA

Abstract

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.

Funder

Connecticut-Storrs Agriculture Experimental Station

USDA National Institute of Food and Agriculture SCRI

Publisher

Oxford University Press (OUP)

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