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High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants

Demirer, Gozde S. and Zhang, Huan and Matos, Juliana L. and Goh, Natalie S. and Cunningham, Francis J. and Sung, Younghun and Chang, Roger and Aditham, Abhishek J. and Chio, Linda and Cho, Myeong-Je and Staskawicz, Brian and Landry, Markita P. (2019) High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants. Nature Nanotechnology, 14 (5). pp. 456-464. ISSN 1748-3387. doi:10.1038/s41565-019-0382-5. https://resolver.caltech.edu/CaltechAUTHORS:20220303-892419000

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Abstract

Genetic engineering of plants is at the core of sustainability efforts, natural product synthesis and crop engineering. The plant cell wall is a barrier that limits the ease and throughput of exogenous biomolecule delivery to plants. Current delivery methods either suffer from host-range limitations, low transformation efficiencies, tissue damage or unavoidable DNA integration into the host genome. Here, we demonstrate efficient diffusion-based biomolecule delivery into intact plants of several species with pristine and chemically functionalized high aspect ratio nanomaterials. Efficient DNA delivery and strong protein expression without transgene integration is accomplished in Nicotiana benthamiana (Nb), Eruca sativa (arugula), Triticum aestivum (wheat) and Gossypium hirsutum (cotton) leaves and arugula protoplasts. We find that nanomaterials not only facilitate biomolecule transport into plant cells but also protect polynucleotides from nuclease degradation. Our work provides a tool for species-independent and passive delivery of genetic material, without transgene integration, into plant cells for diverse biotechnology applications.


Item Type:Article
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https://doi.org/10.1038/s41565-019-0382-5DOIArticle
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ORCID:
AuthorORCID
Demirer, Gozde S.0000-0002-3007-1489
Zhang, Huan0000-0002-1096-4255
Landry, Markita P.0000-0002-5832-8522
Additional Information:© 2019 Nature Publishing Group. Received 08 August 2018. Accepted 18 January 2019. Published 25 February 2019. Issue Date May 2019. The authors acknowledge support from a Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI), a Stanley Fahn PDF Junior Faculty Grant under award no. PF-JFA-1760, a Beckman Foundation Young Investigator Award, a USDA AFRI award, a grant from the Gordon and Betty Moore Foundation, a USDA NIFA award, support from the Chan-Zuckerberg foundation and an FFAR New Innovator Award (to M.P.L). G.S.D. is supported by a Schlumberger Foundation Faculty for the Future Fellowship. L.C. is supported by National Defense Science and Engineering Graduate (NDSEG) Fellowship and by the LAM Foundation. The authors thank C. Gee for assisting with the Imaging-PAM Maxi fluorimeter, A. Schultink and A. Ortega for helpful discussions and C. Jakobson and D. Tullman-Ercek for generously sharing their laboratory resources. The authors also acknowledge support from UC Berkeley Molecular Imaging Center (supported by the Gordon and Betty Moore Foundation), the UC Berkeley Biological Imaging Facility (supported in part by the National Institutes of Health S10 program under award no. 1S10OD018136-01), the QB3 Shared Stem Cell Facility, the Innovative Genomics Institute (IGI), and R. Zalpuri at the Electron Microscopy Lab at UC Berkeley for TEM sample preparation and imaging. Data availability. The data that support the plots within this paper and other findings of this study are available from the corresponding author upon reasonable request. Contributions. G.S.D. and M.P.L. conceived of the project, designed the study and wrote the manuscript. G.S.D. performed the majority of experiments and all data analysis. H.Z. and L.C. performed AFM imaging, and H.Z. also performed nanoparticle internalization experiments into mature leaves and western blot experiments. J.L.M. performed Agrobacterium and wheat transformation experiments. N.S.G. helped with designing ddPCR experiments and performed CNT leaf toxicity confocal imaging and TIRF experiments. F.C. performed nanoparticle internalization experiments into isolated protoplasts. Y.S. performed TEM imaging of leaves. A.J.A. and R.C. prepared the plasmids used in the studies. M.-J.C. performed particle bombardment experiments. All authors have edited and commented on the manuscript, and have given their approval of the final version. The authors declare no competing interests. Journal peer review information: Nature Nanotechnology thanks Neena Mitter, Eleni Stavrinidou and the other anonymous reviewer(s) for their contribution to the peer review of this work.
Funders:
Funding AgencyGrant Number
Burroughs Wellcome FundUNSPECIFIED
Parkinson's FoundationPF-JFA-1760
Arnold and Mabel Beckman FoundationUNSPECIFIED
Department of AgricultureUNSPECIFIED
Gordon and Betty Moore FoundationUNSPECIFIED
Chan-Zuckerberg FoundationUNSPECIFIED
Foundation for Food ResearchUNSPECIFIED
Schlumberger FoundationUNSPECIFIED
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
LAM FoundationUNSPECIFIED
University of California, BerkeleyUNSPECIFIED
NIH1S10OD018136-01
Innovative Genomics InstituteUNSPECIFIED
Issue or Number:5
DOI:10.1038/s41565-019-0382-5
Record Number:CaltechAUTHORS:20220303-892419000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220303-892419000
Official Citation:Demirer, G.S., Zhang, H., Matos, J.L. et al. High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants. Nat. Nanotechnol. 14, 456–464 (2019). https://doi.org/10.1038/s41565-019-0382-5
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113720
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:04 Mar 2022 00:23
Last Modified:04 Mar 2022 00:23

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