Lysozyme-coated nanoparticles for active uptake and delivery of synthetic RNA and plasmid-encoded genes in plants

Abstract

Nanoparticle-mediated delivery of nucleic acids and proteins into intact plants has the potential to modify metabolic pathways and confer desirable traits in crops. Here we show that layered double hydroxide (LDH) nanosheets coated with lysozyme are actively taken up into the root tip, root hairs and lateral root junctions by endocytosis, and translocate via an active membrane trafficking pathway in plants. Lysozyme coating enhanced nanosheet uptake by (1) loosening the plant cell wall and (2) stimulating the expression of endocytosis and other membrane trafficking genes. The lysozyme-coated nanosheets efficiently delivered synthetic mRNA, double-stranded RNA, small interfering RNA and plasmid DNA up to 15 kb in size into tobacco roots, and also functional nucleic acids into leaves, callus, flowers and developing pollen of dicot and monocot species. Thus, lysozyme-coated LDH nanoparticles are a versatile tool for efficiently delivering functional nucleic acids into plants. This work describes protein-coated clay nanoparticles that are actively taken up by roots to deliver synthetic RNA and DNA-encoded genes into plants. The nanoparticles can also deliver functional nucleic acids directly into other plant tissues.