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Proc Natl Acad Sci U S A. 2019 Apr 9;116(15):7543-7548. doi: 10.1073/pnas.1818290116. Epub 2019 Mar 25.

DNA nanostructures coordinate gene silencing in mature plants.

Author information

1
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720.
2
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.
3
Division of Physical Biology & Bioimaging Center, Shanghai Synchrotron Radiation Facility, Chinese Academy of Sciences Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China.
4
School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.
5
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720; landry@berkeley.edu.
6
Innovative Genomics Institute, Berkeley, CA 94720.
7
California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, CA 94720.
8
Chan-Zuckerberg Biohub, San Francisco, CA 94158.

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.

KEYWORDS:

DNA nanotechnology; DNA origami; RNA delivery; plant biotechnology; siRNA gene silencing

PMID:
30910954
PMCID:
PMC6462094
[Available on 2019-09-25]
DOI:
10.1073/pnas.1818290116
[Indexed for MEDLINE]

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