Poster Presentation International Plant Molecular Biology Conference 2024

Nanoclay-delivered mobile RNA for applications in plant biotechnology (#44)

Wang Xu 1 , Jiaxi Yong 2 , Shengnan Bi 1 , Anne Sawyer 1 3 , Neena Mitter 3 , Zhiping Xu 2 , Friedrich Kragler 4 , Bernard Carroll 1
  1. School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QUEENSLAND, Australia
  2. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia
  3. Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, Brisbane, Queensland, Australia
  4. Max Planck Institute of Molecular Plant Physiology, Wissenschaftspark Golm, Am Muehlenberg, German

Genetic improvement of crops relies on traditional plant breeding, as well as genetic modification for a limited number of crop species. However, these processes are lengthy and public acceptance of genetically modified (GM) crops is faltering. The CRISPR gene editing system has enormous potential as a tool for crop improvement, but currently requires genetic modification. However, heritable gene editing in Arabidopsis has been achieved by grafting non-GM shoots onto GM rootstocks that express both the CRISPR mRNA and guide RNAs fused to a graft-transmissible tRNA motif (Yang et al., 2023).

Layered double hydroxide (LDH) nanosheets up to 50 nm in diameter traverse the plant cell wall and facilitate the uptake of functional RNA into intact plants (Yong et al., 2021, 2022). Furthermore, we have recently discovered that protein coating of the LDH nanosheets greatly enhances their uptake by plant roots via endocytosis and the membrane trafficking pathway. We have used the protein-coated LDH nanosheets to not only transiently deliver transgenes but to deliver functional mRNA into intact plants for the first time (Yong et al., in prep). Furthermore, we have shown that nanoparticle delivery of a transgene encoding mRNA fused to a graft-transmissible tRNA motif into plant roots enables the systemic movement of the mRNA and the expression of the encoded protein in the shoots. The study contributes valuable insights into the potential application of nanotechnology and mobile RNA in plant biotechnology, offering the possibility of non-GM trait modification in intact plants of both model and crop species.

  1. YONG, J., WU, M., ZHANG, R., BI, S., MANN, C. W. G., MITTER, N., CARROLL, B. J. & XU, Z. P. 2022. Clay nanoparticles efficiently deliver small interfering RNA to intact plant leaf cells. Plant Physiol, 190, 2187-2202
  2. YONG, J., ZHANG, R., BI, S., LI, P., SUN, L., MITTER, N., CARROLL, B. J. & XU, Z. P. 2021. Sheetlike clay nanoparticles deliver RNA into developing pollen to efficiently silence a target gene. Plant Physiol, 187, 886-899.
  3. YANG, L., MACHIN, F., WANG, S., SAPLAOURA, E. & KRAGLER, F. 2023a. Heritable transgenefree genome editing in plants by grafting of wild-type shoots to transgenic donor rootstocks. Nat Biotechnol, 41, 958-967.