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DNA nanostructures coordinate gene silencing in mature plants

Zhang, Huan and Demirer, Gozde S. and Zhang, Honglu and Ye, Tianzheng and Goh, Natalie S. and Aditham, Abhishek J. and Cunningham, Francis J. and Fan, Chunhai and Landry, Markita P. (2019) DNA nanostructures coordinate gene silencing in mature plants. Proceedings of the National Academy of Sciences of the United States of America, 116 (15). pp. 7543-7548. ISSN 0027-8424. PMCID PMC6462094. doi:10.1073/pnas.1818290116. https://resolver.caltech.edu/CaltechAUTHORS:20220302-323665000

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Abstract

Delivery of biomolecules to plants relies on Agrobacterium infection or biolistic particle delivery, the former of which is amenable only to DNA delivery. The difficulty in delivering functional biomolecules such as RNA to plant cells is due to the plant cell wall, which is absent in mammalian cells and poses the dominant physical barrier to biomolecule delivery in plants. DNA nanostructure-mediated biomolecule delivery is an effective strategy to deliver cargoes across the lipid bilayer of mammalian cells; however, nanoparticle-mediated delivery without external mechanical aid remains unexplored for biomolecule delivery across the cell wall in plants. Herein, we report a systematic assessment of different DNA nanostructures for their ability to internalize into cells of mature plants, deliver siRNAs, and effectively silence a constitutively expressed gene in Nicotiana benthamiana leaves. We show that nanostructure internalization into plant cells and corresponding gene silencing efficiency depends on the DNA nanostructure size, shape, compactness, stiffness, and location of the siRNA attachment locus on the nanostructure. We further confirm that the internalization efficiency of DNA nanostructures correlates with their respective gene silencing efficiencies but that the endogenous gene silencing pathway depends on the siRNA attachment locus. Our work establishes the feasibility of biomolecule delivery to plants with DNA nanostructures and both details the design parameters of importance for plant cell internalization and also assesses the impact of DNA nanostructure geometry for gene silencing mechanisms.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1073/pnas.1818290116DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc6462094/PubMed CentralArticle
ORCID:
AuthorORCID
Zhang, Huan0000-0002-1096-4255
Demirer, Gozde S.0000-0002-3007-1489
Landry, Markita P.0000-0002-5832-8522
Additional Information:© 2019. Published under the PNAS license. Edited by Catherine J. Murphy, University of Illinois at Urbana–Champaign, Urbana, IL, and approved February 27, 2019 (received for review October 27, 2018) This article is a PNAS Direct Submission. We acknowledge the support of the Burroughs Wellcome Fund (Career Awards at the Scientific Interface), a US Department of Agriculture (USDA) Agriculture and Food Research Initiative grant with Award 2018-67021-27964, a Foundation for Food and Agriculture Research New Innovator Award, an NSF-USDA-Biotechnology and Biological Sciences Research Council Grant, and the Berkeley Molecular Imaging Center and QB3 Shared Stem Cell facilities. H.Z. acknowledges the National Natural Science Foundation of China (Grant 21605153). G.S.D. is supported by the Schlumberger Foundation. Author contributions: Huan Zhang, G.S.D., Honglu Zhang, N.S.G., A.J.A., F.J.C., and M.P.L. designed research; Huan Zhang, G.S.D., Honglu Zhang, T.Y., N.S.G., A.J.A., and F.J.C. performed research; Honglu Zhang and C.F. contributed new reagents/analytic tools; Huan Zhang, G.S.D., T.Y., N.S.G., A.J.A., F.J.C., and M.P.L. analyzed data; and Huan Zhang, G.S.D., and M.P.L. wrote the paper. Huan Zhang, G.S.D., and Honglu Zhang contributed equally to this work. The authors declare no conflict of interest
Funders:
Funding AgencyGrant Number
Burroughs Wellcome FundUNSPECIFIED
Department of Agriculture2018-67021-27964
Foundation for Food ResearchUNSPECIFIED
NSFUNSPECIFIED
Biotechnology and Biological Sciences Research Council (BBSRC)UNSPECIFIED
National Natural Science Foundation of China21605153
Schlumberger FoundationUNSPECIFIED
Subject Keywords:plant biotechnology | DNA nanotechnology | siRNA gene silencing | RNA delivery | DNA origami
Issue or Number:15
PubMed Central ID:PMC6462094
DOI:10.1073/pnas.1818290116
Record Number:CaltechAUTHORS:20220302-323665000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220302-323665000
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113688
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:03 Mar 2022 23:49
Last Modified:03 Mar 2022 23:49

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