Nanoneedle-enabled platform for precision integration of large DNA into crop genomes

Overview

This silicon nanoneedle–based genome engineering platform addresses the delivery bottleneck that limits precise integration of large DNA sequences into crop genomes. Conventional plant transformation methods struggle to move large DNA molecules intact across the cell wall, restricting trait insertion. Nanoneedles physically penetrate plant cells with minimal damage, allowing the co-delivery of intact large DNA donor templates alongside genome-editing components — including CRISPR-Cas machinery — improving intracellular access for a wide range of cargo types.

The platform has delivered reporter plasmids, viral vectors, gene-silencing reagents, and CRISPR-Cas9 across several plant species and target tissues, including fluorescent reporter plots as large as 14 kb and viral vectors as large as 20 kb. This establishes a strong technical foundation for programmable, precise gene insertion in commercially important crops — a key capability for advanced breeding and trait stacking.

Technical specifications

How it works:

• Silicon nanoneedles penetrate plant cell boundaries directly, eliminating the need for specialized biological carriers and enabling transfer of very large molecules.
• Co-delivery of multiple payloads — including donor DNA and editing agents — is achieved in a single physical exposure.

Demonstrated capabilities:

• Delivery of an 11 kb GFP plasmid and 14 kb RUBY plasmid with confirmed functional expression in Arabidopsis.
• Co-delivery of 20 kb TRV1 and 9 kb TRV2 viral vectors in tobacco, with functional gene silencing confirmed by PDS and chlorophyll assays.
• CRISPR-Cas9 delivery to kailan hypocotyls, regenerating whole plants with sequence-validated BRC1 and AOP2 gene knockouts.
• Proven cargo compatibility across Arabidopsis, tobacco, choysum pollen, and kailan tissues, with minimal cell damage.

Development path:

• Combine the nanoneedle delivery platform with Cas12a targeting and optimized donor/repair strategies to achieve predictable 7–10 kb site-specific integration.
• Expand payload capacity toward 25 kb to support complex trait or gene-stacking bundles.
• Validate the system in regenerable tissue of additional crop backgrounds, including soybean and oilseed rape.

Technology readiness level

The platform is at technology readiness level 3–5 (laboratory/bench validation). Functional delivery and genome editing have been confirmed in multiple plant species, including sequence-validated knockout regeneration. Current research is extending the platform to additional crop species and target-defined integration, followed by validation of integration, fidelity, stable inheritance, and reproducibility — milestones required before field-level or commercial deployment.