Nanoparticle-Mediated Transgene Expression and Silencing in Agriculturally-Relevant Plants

Authors: 
Demirer, G. S., University of California
Zhang, H., University of California
Matos, J., University of California
Chio, L., University of California
Staskawicz, B., University of California
Landry, M., Chan Zuckerberg Biohub

Food security has been threatened with decreasing crop yields and increasing consumption in the light of population growth and climate change. Plant biotechnology is critical to address the world’s leading challenges in meeting our growing food and energy demands, and as a tool for scalable pharmaceutical manufacturing. Over the past several decades, remarkable progress has been made in biotechnology with the improvement of genome editing and sequencing tools. However, of the many biological systems and organisms in which transgenic biotechnologies have been implemented, plants are vastly underrepresented. The challenge of gene delivery to plants is attributed to a transport limitation: the presence of the multilayered and rigid plant cell wall, otherwise absent in animal cells1. To date, plant biotechnology lacks a method that allows passive delivery of diverse biomolecules into a broad range of plant species without the aid of external force.

Herein, we develop a nanoparticle-based platform that can deliver functional biomolecules into both model and crop plants with high efficiency and no toxicity2. Carbon nanotubes (CNTs) are leveraged to deliver DNA into mature arugula (dicot) and wheat (monocot) leaves, among others, from which we obtain strong transient protein expression. Additionally, we achieve 95% transient gene silencing in Nicotiana benthamiana leaves through CNT-mediated delivery of small interfering RNA. Furthermore, we discuss the implementation of DNA origami techniques for elucidating the mechanism of nanoparticle transport in the plant. This study establishes efficient transient gene expression and silencing in mature plant leaves through passive nanoparticle-mediated delivery of functional biomolecules and can enable multiplexable and high-throughput genetic plant transformations.

  1. Cunningham, F.J., Goh N.S. and Demirer, G.S. et al. (2018) Nanoparticle-mediated delivery towards advancing plant genetic engineering. Trends Biotechnol.
  2. Demirer, G.S. et al. (2018) High Aspect Ratio Nanomaterials Enable Biomolecule Delivery and Transgene Expression or Silencing in Mature Plants. bioRxiv DOI: 10.1101/179549