Nanohydroxyapatite-based nanoparticle delivery system for gene-edited plant production

Overview

This research proposes a nanohydroxyapatite (nHA) rod-based delivery system for plant genetic engineering and gene editing. Conventional plant transformation methods often rely on Agrobacterium or biolistics, which can be costly, time-consuming, and limited by species compatibility. The proposed approach leverages nHA rods as nanocarriers that are small enough to traverse plant cell walls, biodegradable, and biocompatible, enabling direct delivery of molecular cargo such as plasmid DNA into plant tissues. The goal is to combine this nanobiotechnology with de novo meristem regeneration and genome editing to produce gene-edited plants more efficiently.

Technical specifications

Key features:

• nHA rods approximately 10 nm in diameter, within the 5–20 nm size exclusion limit of plant cell walls
• Plasmid DNA attaches to nHA surfaces via electrostatic interactions, forming stable conjugates
• Partial protection of delivered DNA from nuclease degradation in planta
• Successful transient expression of reporter genes (GUS and GFP) in multiple plant species and tissues, including leaf and seed
• Planned modifications: increased surface area, reduced diameter to 5 nm, and short peptide tags to mediate nuclear transport of gene-editing cargo
• Validation strategy includes knockout of an albino gene in tobacco for visual identification of edited plantlets, confirmed by PCR and sequencing

Technology readiness level

The technology is currently at an early-to-mid stage of development. Transient gene expression has been demonstrated in multiple plant species, confirming that nHA-pDNA conjugates can deliver molecular cargo into plant tissues. The next phase focuses on optimizing the nanoparticles for larger gene-editing cargo loads, incorporating nuclear-targeting peptides, and producing stable gene-edited plantlets. The estimated timeline for this validation phase is approximately one year. Long-term translation targets major crops including canola, soybean, and wheat.