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The DNA–carbon nanotube binding mode determines the efficiency of carbon nanotube-mediated DNA delivery to intact plants

Ali, Zahir and Serag, Maged F. and Demirer, Gozde S. and Torre, Bruno and di Fabrizio, Enzo and Landry, Markita P. and Habuchi, Satoshi and Mahfouz, Magdy M. (2022) The DNA–carbon nanotube binding mode determines the efficiency of carbon nanotube-mediated DNA delivery to intact plants. . (Unpublished)

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Efficient delivery of DNA, RNA, and genome engineering machinery to plant cells will enable efforts to genetically modify plants for global food security, sustainable energy production, synthetic biology applications, and climate change resilience. For the delivery of functional genetic units into plant cells, charged nanoparticles, particularly carbon nanotubes (CNT), have attracted considerable interest. Although some success has been achieved using CNT-based approaches, the efficiency, batch reproducibility, and the limits of their applicability remain to be assessed. Here, we provide a mechanistic understanding of plasmid DNA-loaded CNTs based transfection of plant cells, and factors affecting the expression of the transformed plasmid. We show that transfection is inherently limited by the presence of the cell wall, Coulomb interactions between DNA and polymer coated CNT, and DNA size, whereas expression of the transformed plasmid is limited by relative gene-to-plasmid size and the intracellular accessibility of DNA. We further show that the formation of partially condensed DNA on the CNT surface is a prerequisite for successful transfection and expression. Furthermore, DNA does not detach completely from the CNT, so the accessibility of the transcription machinery to DNA is the key for transformation efficiency. This irreversible DNA plasmid binding and partial condensation limit the length of DNA that can be expressed, thus negatively affecting efficiency and reproducibility. Understanding the underlying mechanisms and limitations of CNT-mediated delivery of DNA through the plant cell wall is of considerable importance in guiding efforts to design nanomaterials for efficient transformation, trait engineering, and synthetic biology applications.

Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
Demirer, Gozde S.0000-0002-3007-1489
Landry, Markita P.0000-0002-5832-8522
Mahfouz, Magdy M.0000-0002-0616-6365
Additional Information:The content is available under CC BY 4.0 License We would like to thank members of the genome engineering and synthetic biology laboratory for insightful discussions and technical support. This work was supported, by KAUST grant to M. Mahfouz. Author Contributions. MM, SH and ML conceived research. ZA and MS designed the research. ZA, MS, GD, and BT performed the research. ZA, MS, MM, SH wrote the paper with input from ML, GD, MS, BT and EF. The authors declare no competing financial interests.
Funding AgencyGrant Number
King Abdullah University of Science and Technology (KAUST)UNSPECIFIED
Subject Keywords:carbon nanotubes, DNA, plant cell, gene delivery, genetic transformation, plants genome engineering, nanobiotechnology
Record Number:CaltechAUTHORS:20220302-323858000
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Official Citation:Ali Z, Serag M, Demirer G, et al. The DNA–carbon nanotube binding mode determines the efficiency of carbon nanotube-mediated DNA delivery to intact plants . ChemRxiv. 2021. doi:10.26434/chemrxiv-2021-43vr3
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113699
Deposited By: George Porter
Deposited On:03 Mar 2022 18:32
Last Modified:03 Mar 2022 18:32

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