Large and combinatory biological cargo delivery for plant cell genome engineering

Overview

This plant cell delivery technology introduces DNA, proteins, and RNA in a single step, with the capacity to handle payloads in the micrometer range. Unlike conventional transformation methods, which are limited by cargo size, this approach delivers material directly into the cytoplasm. That opens the way for engineered, large, contiguous DNA segments to be inserted home in their target locus.

The key advantage is that the editing machinery does not need to be inserted into the genome. Because the editing components and donor template are co-delivered without genomic integration, the process achieves transgene-free insertion — a valuable outcome for generating crops with a clean genetic background and for downstream regulatory and product development.

The technology is positioned for early genome engineering programs, where large insertions or multi-component edits are needed but are currently limited by cargo capacity. Applications include inserting large gene cassettes for trait engineering, customizing plant metabolic pathways, and creating defined single-step edits in crop species.

Technical specifications

Key features:

• Co-delivery of DNA, proteins, and RNA in a single cytoplasmic delivery event
• Handles micrometer-scale payloads, beyond the limit of many existing plant transformation tools
• Direct delivery of the editing machinery and donor template enables large targeted insertions without prior integration of the editing tools
• Transgene-free workflow with editing machinery only transiently present
• Concept parallels mammalian microinjection studies, where direct cytoplasmic delivery of CRISPR-Cas and a donor template achieved roughly 40% insertion of fragments as large as 7 kb

Validation pipeline:

• Demonstrate large targeted insertion in potato to verify the feasibility of the combined delivery method (~6 months)
• Concurrently develop regeneration protocols for soybean (6–12 months)
• Following the protocol milestones, validate the delivery in soybean for targeted insertion of a custom gene of interest (post-protocol, ~6 months)
• Finalize with a product generation step, inserting a transgene of interest in the target locus to generate a first polished product

Technology readiness level

The technology is at the early-to-validated research stage. The underlying principle of combined cytoplasmic delivery of multiple biological cargoes has been demonstrated, but the targeted large-insertion capability is still being validated. The first feasibility checkpoint in potato is ongoing in the near term (around 6 months), with soybean validation expected after regeneration protocols are in place (approximately 18–24 months in total). The method is being prepared for a first functional product, but its widespread validation in target crop species remains to be completed.