DNA Nanotechnology Enabled Cargo Delivery in Plants - DNA Nanotechnology Enabled Cargo Delivery in Plants

ABSTRACT

Plant bioengineering has great potential to generate high yielding and pathogen-resistant crops, and is at the core of sustainability efforts. However, unlike mammalian cells, plant cells have a cell wall which poses the dominant barrier to exogenous biomolecule delivery. To systematically assess the factor for efficient internalization into plant cells, we designed and synthesized a series of DNA nanostructures of controllable size, shape, stiffness, and compactness, and designed attachment loci onto which small interfering RNA (siRNA) was conjugated. We found that DNA nanostructures can passively internalize into plant cells and deliver siRNA to mature plant tissues. We showed that the DNA nanostructure size, shape, compactness, stiffness, and location of the siRNA attachment locus on the nanostructure affect nanostructure internalization into plant cells and the corresponding gene silencing efficiency. Furthermore, with the rapidly growing interest in nanoparticle-mediated delivery of DNA and RNA to plants, it requires a better understanding of how nanoparticles and their cargoes translocate in plant tissues and into plant cells. With the help of DNA technology, we design a model nanoparticle system based on DNA-functionalized gold nanoparticles (AuNP) of varied sizes and shapes, then we utilize confocal microscopy and transmission electron microscopy (TEM) to track AuNP movement fate following infiltration into leaves, and directly visualize NP interactions with plant cells. Our results indicate the importance of nanoparticle size in efficient biomolecule delivery, and highlight nanoparticle features of importance for transport within plant tissues, providing a mechanistic overview of how nanoparticles can be designed to achieve efficacious bio-cargo delivery for future developments in plant nanobiotechnology.