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Title: DNA nanostructures coordinate gene silencing in mature plants

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 plantsmore » 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.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2]; ORCiD logo [3]
  1. Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engineering
  2. Chinese Academy of Sciences (CAS), Shanghai (China). Shanghai Inst. of Applied Physics, Key Lab. of Interfacial Physics and Technology, Division of Physical Biology & Bioimaging Center, Shanghai Synchrotron Radiation Facility; Shanghai Jiao Tong Univ. (China). School of Chemistry and Chemical Engineering
  3. Univ. of California, Berkeley, CA (United States). Dept. of Chemical and Biomolecular Engineering; Innovative Genomics Inst., Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences; Chan-Zuckerberg Biohub, San Francisco, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1561909
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 116; Journal Issue: 15; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Zhang, Huan, Demirer, Gozde S., Zhang, Honglu, Ye, Tianzheng, Goh, Natalie S., Aditham, Abhishek J., Cunningham, Francis J., Fan, Chunhai, and Landry, Markita P. DNA nanostructures coordinate gene silencing in mature plants. United States: N. p., 2019. Web. doi:10.1073/pnas.1818290116.
Zhang, Huan, Demirer, Gozde S., Zhang, Honglu, Ye, Tianzheng, Goh, Natalie S., Aditham, Abhishek J., Cunningham, Francis J., Fan, Chunhai, & Landry, Markita P. DNA nanostructures coordinate gene silencing in mature plants. United States. doi:10.1073/pnas.1818290116.
Zhang, Huan, Demirer, Gozde S., Zhang, Honglu, Ye, Tianzheng, Goh, Natalie S., Aditham, Abhishek J., Cunningham, Francis J., Fan, Chunhai, and Landry, Markita P. Mon . "DNA nanostructures coordinate gene silencing in mature plants". United States. doi:10.1073/pnas.1818290116. https://www.osti.gov/servlets/purl/1561909.
@article{osti_1561909,
title = {DNA nanostructures coordinate gene silencing in mature plants},
author = {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.},
abstractNote = {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.},
doi = {10.1073/pnas.1818290116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 15,
volume = 116,
place = {United States},
year = {2019},
month = {3}
}

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