Targeted transformation platform for predictable large-fragment genome insertion in commercial crops

Overview

This robust targeted transformation platform addresses a major bottleneck in crop genome engineering: the predictable introduction of large DNA fragments at predefined genomic loci. It combines precision genome engineering technologies, including zinc finger nucleases (ZFNs) and CRISPR/Cas systems, with PEG-mediated protoplast transformation in canola. The platform is designed to enable efficient gene targeting and site-specific DNA integration, resulting in predictable and consistent transgene expression while minimizing insertional effects. It supports precise trait deployment and trait stacking, which can simplify product development, breeding workflows, and downstream regulatory characterization for commercial crops.

Technical specifications

The platform integrates three key components: efficient gene delivery, precision gene-targeting reagents with optimized donor DNA delivery, and recovery of on-target events.

Key features:

• Uses PEG-mediated protoplast transfection and regeneration in canola as a rapid system for optimizing donor design, repair-pathway modulation, and targeted integration efficiency. Protoplasts—plant cells with their cell walls removed—allow quick testing of DNA-delivery and editing configurations.
• Recovers stable transformants with precise, single-copy insertions at defined genomic loci.
• Designed for minimal off-target effects and stable inheritance.
• Compatible with Agrobacterium-mediated transformation workflows in canola and soybean, supporting commercial trait development and trait-stacking applications.

Technology readiness level

This technology is in an early development and validation phase. The initial workstream will assess editing reagents, donor templates, and enabling technologies before incorporating them into the genome integration workflow. These components will be tested on the PEG-mediated canola protoplast platform to optimize donor design, repair pathway modulation, and targeted insertion efficiency. Promising configurations will then be integrated into the full pipeline, validated for targeted integration of large fragments, molecularly characterized, and adapted to Agrobacterium-mediated transformation in canola and soybean to demonstrate transferability, stable inheritance, and scalability.